DNA/RNA Amplification Overview AccuPower PreMix series 는세계적으로기술력을인정받은특허기술로보다경제적인가격과편리한방법으로실험할수있는제품입니다. Conventional PCR, Real-Time PCR 수행을위한 DNA ampli

DNA Amplification RNA Amplification Real-Time PCR Customized PCR DNA/RNA Amplification Phone: 1588-9788 (ext.4->2) Email: accupower-support @bioneer.co.kr

DNA/RNA Amplification Overview AccuPower PreMix series 는세계적으로기술력을인정받은특허기술로보다경제적인가격과편리한방법으로실험할수있는제품입니다. Conventional PCR, Real-Time PCR 수행을위한 DNA amplification kit 과 reverse transcription 을통해 cdna 합성이가능한 RNA amplification kit 는 life science 연구에필요한 total solution 을제공합니다. AccuPower PreMix series 는 enzyme, dntps 및 reaction buffer 등 PCR 수행에필요한구성성분을혼합하여 1 회분량씩진공건조시킨제품입니다. 실험자는 template DNA, primers 및 D.W. 만첨가함으로써편리하고정확하게실험할수있습니다. AccuPower PreMix series 의모든제품은 PCR 반응혼합액에안정화제가첨가되어상온에서도안정하게실험을할수있으며 ice 에서실험하는번거로움을최소화하였습니다. 상온 (25 ) 보관할경우에는한달동안, 냉동보관할경우에는 2 년동안활성이안정적으로유지됩니다. 또한, 전기영동에필요한 tracking dye 와비중증가제가포함되어있기때문에반응액을그대로 agarose gel 전기영동에사용할수있어서편리합니다. One-batch system 으로대량생산하고, 엄격한 ISO 9001 품질규격에의한품질검사를통해최상의실험결과와높은재현성을보장합니다. Selection Guide of AccuPower PreMix Series DNA Amplification Applications Products Standard PCR HotStart PCR Prevent Carryover Contamination PCR for Gene Cloning PCR for TA Cloning High Fidelity PCR Long Range PCR GC Rich PCR Multiplex PCR AccuPower PCR AccuPower PCR (with UDG) AccuPower Taq PCR AccuPower HotStart PCR AccuPower HotStart PCR (with UDG) AccuPower GoldHotStart Taq PCR AccuPower PyroHotStart Taq PCR AccuPower HotStart Pfu PCR AccuPower ProFi Taq PCR AccuPower Pfu PCR AccuPower Multiplex PCR AccuPower Gold Multiplex PCR 2

DNA/RNA Amplification RNA Amplification cdna Synthesis Kits Applications Products Standard RT High Efficiency RT RT of Secondary Structured RNA HotStart RT AccuPower RT AccuPower CycleScript RT AccuPower RocketScript TM RT AccuPower RocketScript TM Cycle RT AccuPower RocketScript TM RT (RNase H-) One-step RT-PCR Kits Applications Products Standard RT- PCR RT-PCR of Secondary Structured RNA High Specificity & High Sensitivity HotStart RT Prevent Carryover Contamination Multiplex RT-PCR AccuPower RT-PCR AccuPower RocketScript TM RT-PCR AccuPower Dual-HotStart TM RT-PCR AccuPower Dual-HotStart TM RT-PCR (with UDG) AccuPower RocketPlex RT-PCR Real-Time PCR Products AccuPower GreenStar TM qpcr AccuPower DualStar TM qpcr AccuPower Plus DualStar TM qpcr AccuPower Plus DualStar TM qpcr (with UDG) AccuPower Dual-HotStart TM RTqPCR HotStart RT HotStart qpcr Applications qpcr (Intercalating dye) qpcr (TaqMan Probe) Prevent Carryover Contamination www.bioneer.co.kr 3

DNA/RNA Amplification Specifications of AccuPower Premix Series PCR & One-step RT-PCR Product Product Size 5 ->3 Exonuclease 3 ->5 Exonuclease Specificity Fidelity GC-rich Leaves 3'-A PCR 10 kb No No *** *** *** Yes PCR (with UDG) 10 kb No No *** *** *** Yes Taq PCR 10 kb Yes No *** *** *** Yes Pfu PCR 10 kb No Yes **** ***** *** No ProFi Taq PCR 30 kb Yes Yes **** **** *** Yes HotStart PCR 10 kb No No **** *** **** Yes HotStart PCR (with UDG) 10 kb No No **** *** **** Yes GoldHotStart Taq PCR 10 kb Yes No **** *** **** Yes PyroHotStart Taq PCR 10 kb Yes No **** *** **** Yes HotStart Pfu PCR 15 kb No Yes **** ***** *** No Multiplex PCR 1~20 frag. No No **** *** *** Yes Gold Muliplex PCR 1~20 frag. No No **** *** *** Yes RT-PCR 6 kb No No - - *** Yes RocketScript TM RT-PCR 6 kb No No - - *** Yes RocketScript TM RT-PCR (RNase H-) 9 kb Yes No - - *** Yes Dual-HotStart TM RT-PCR 3kb Yes No ***** - **** Yes Dual-HotStart TM RT-PCR (with UDG) 3kb Yes No ***** - **** Yes RocketPlex RT-PCR 1 kb No No **** - *** Yes Reverse Transcription Product Product Size RNase H Activity DNase Activity RNase Activity GC-rich RT 9 kb Yes No No *** CycleScript RT 9 kb Yes No No *** RocketScript TM RT 10 kb Yes No No *** RocketScript TM Cycle RT 10 kb Yes No No *** RocketScript TM RT (RNase H-) 12.5 kb No No No **** qpcr & One-step RT-qPCR Product 5 ->3 Exonuclease 3 ->5 Exonuclease Specificity GC-rich Leaves 3'-A GreenStar TM qpcr No No **** **** Yes Dual-Star TM qpcr Yes No ***** **** Yes Plus Dual-Star TM qpcr Yes No **** **** Yes Plus Dual-Star TM qpcr (with UDG) Yes No **** **** Yes Dual-HotStart TM RT-qPCR Yes No ***** **** Yes 4

DNA/RNA Amplification Selection Guide of Enzymes Product DNA Polymerase HotStart DNA Polymerase Reverse Transcriptase Top Taq Standard PCR HotStart PCR PCR for Gene Cloning Pfu PCR Long Range PCR ProFi Taq Applications GC Rich PCR Multiplex PCR Top Taq M-MLV Standard RT RT Cyclic RT CycleScript RocketScript TM RocketScript TM RNase H Minus Long range RT Specifications of Enzymes Product Product Size 5 ->3 Exonuclease 3 ->5 Exonuclease Specificity Fidelity GC-rich Leaves 3'-A Top DNA Polymerase 10 kb No No **** *** *** Yes Taq DNA Polymerase 10 kb Yes No **** *** *** Yes HotStart Taq DNA Polymerase 12 kb Yes No **** *** **** Yes HotStart DNA Polymerase 12 kb No No **** *** **** Yes ProFi Taq DNA Polymerase 30 kb Yes Yes **** *** **** Yes Pfu DNA Polymerase 10 kb No Yes *** **** ** No M-MLV Reverse Transcriptase 9 kb - - - - *** - CycleScript Reverse Transcriptase 9 kb - - - - *** - RocketScript TM Reverse Transcriptase 10 kb - - - - *** - RocketScript TM RTase RNase H(-) 12.5 kb - - *** - www.bioneer.co.kr 5

01 DNA Amplification Standard PCR Kit AccuPower PCR PreMix 7 AccuPower PCR PreMix (with UDG) 9 AccuPower Taq PCR PreMix 11 HotStart PCR Kit AccuPower HotStart PCR PreMix 13 AccuPower HotStart PCR PreMix (with UDG) 15 AccuPower GoldHotStart Taq PCR PreMix New 17 AccuPower PyroHotStart Taq PCR PreMix 19 AccuPower HotStart Pfu PCR PreMix 21 High Fidelity PCR Kit AccuPower Pfu PCR PreMix 23 Long & High Fidelity PCR Kit AccuPower ProFi Taq PCR PreMix 25 Multiplex PCR Kit AccuPower Multiplex PCR PreMix 27 AccuPower Gold Muliplex PCR PreMix 29 DNA Polymerase Top DNA Polymerase 31 Taq DNA Polymerase 33 Pfu DNA Polymerase 35 ProFi Taq DNA polymerase 37 HotStart DNA Polymerase 39 HotStart Taq DNA Polymerase 41 Conventional PCR Instrument AllInOneCycler TM Go to M. Instruments

AccuPower PCR PreMix 동결건조 PCR 제품의 Global Standard Specifications Enzyme: Top DNA polymerase 5 to 3 exonuclease activity: No 3 to 5 exonuclease activity: No 3 - A overhang: Yes Fragment size: ~ 10 kb Application Conventional PCR Primer extension TA cloning Gene sequencing Description AccuPower PCR PreMix 는바이오니아의특허기술로 Top DNA polymerase, dntps, reaction buffer 등 PCR 수행에필요한구성성분을혼합하여 1 회분량씩동결건조시킨제품입니다. 안정화물질이첨가되어있어상온 (25 ) 보관할경우에는한달동안, 냉동보관할경우에는 2 년동안활성이안정적으로유지되는획기적인제품입니다. Features and Benefits Stability: AccuPower PCR PreMix 는 PCR 반응혼합액에안정화제를첨가하여건조시킨제품으로열안전성이떨어지는 solution type 제품의단점을보완하였으며, 장기간보관하더라도활성을안정적으로유지합니다. 또한 95, 90 분열처리에도효소활성이안정적으로유지되기때문에반복적인 cycle 에도증폭효율이높습니다. Ease-of-use: 각각의 PCR tube 에 DNA polymerase 와 PCR 반응에필요한모든구성성분이포함되어있어 template DNA, primer set, D.W. 만넣어바로 PCR 반응을수행할수있습니다. 또한전기영동시에필요한 tracking dye 와침강제가포함되어있어별도로 sample loading buffer 를첨가할필요가없으므로간편하게사용할수있습니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. Storage Temperature -20 Experimental Data I II III IV M 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 Figure 1. Comparison of sensitivity test for PCR PreMix and other companies products using serial diluted human gdna. Amplification of the human insulin receptor gene. I: AccuPower PCR PreMix II: A company Taq DNA polymerase III: B company Taq DNA polymerase IV: C company PCR PreMix Lane 1: Human gdna 10 ng Lane 2: Human gdna 1 ng Lane 3: Human gdna 100 pg Lane 4: Human gdna 10 pg M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030) Sensitivity: 뛰어난민감도와증폭효율로미량의 human gdna target 을검출합니다. 타사제품과의비교실험을통해우수한민감도를확인했습니다. www.bioneer.co.kr 7

AccuPower PCR PreMix A 1 2 3 4 B 1 2 3 4 C 1 2 3 4 M Figure 2. Comparison of processivity test for AccuPower PCR PreMix and other companies products using lambda DNA. Rxn. condition 95 5 min, [94 30 sec, 57 30 sec, 72 30 sec] (30 cycles), 72 5 min. A: AccuPower PCR PreMix B: S company s Taq premix type C: T company s Taq premix type Lane 1: 1 kb fragment of Lambda DNA Lane 2: 2 kb fragment of Lambda DNA Lane 3: 3 kb fragment of Lambda DNA Lane 4: 4 kb fragment of Lambda DNA M: 100 bp plus DNA Ladder (Bioneer, Cat. no. D-1035) Figure 3. Comparison of thermostability of AccuPower PCR PreMix. AccuPower PCR PreMix is incubated at 95 with various time. A: 30 min, B: 60 min, C: 90 min Lane 1: Human genomic DNA 10 ng Lane 2: Human genomic DNA 1 ng Lane 3: Human genomic DNA 100 pg Lane 4: Human genomic DNA 10 pg M: 100 bp plus DNA ladder (Bioneer, Cat. no. D-1035) Ordering Information Cat. no. Product Description K-2011 AccuPower PCR PreMix, 0.5 ml thin-wall microtube / 100 tubes, 50 μl rxn K-2012 AccuPower PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 20 μl rxn K-2013 AccuPower PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 50 μl rxn K-2016 AccuPower PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 20 μl rxn K-2017 AccuPower PCR PreMix, 0.2 ml thin-wall tubes with individual cap / 480 tubes, 50 μl rxn K-2018 AccuPower PCR Master Mix, 1 ml of 2X master mix solution K-2018-1 AccuPower PCR Master Mix, 10 ml of 2X master mix solution K-2080-1 AccuPower PCR PreMix, thin-wall 384-well full-skirted plate, 5 μl rxn K-2080-2 AccuPower PCR PreMix, thin-wall 384-well full-skirted plate, 10 μl rxn K-2080-3 AccuPower PCR PreMix, thin-wall 384-well full-skirted plate, 20 μl rxn K-2260-1 AccuPower PCR PreMix, thin-wall 96-well flat plate, 10 μl rxn K-2260-2 AccuPower PCR PreMix, thin-wall 96-well full-skirted plate, 10 μl rxn K-2260-3 AccuPower PCR PreMix, thin-wall 96-well semi-skirted plate, 10 μl rxn K-2260-4 AccuPower PCR PreMix, thin-wall 96-well flat plate, 20 μl rxn K-2260-5 AccuPower PCR PreMix, thin-wall 96-well full-skirted plate, 20 μl rxn K-2260-6 AccuPower PCR PreMix, thin-wall 96-well semi-skirted plate, 20 μl rxn 8

AccuPower PCR PreMix (with UDG) 증폭된 PCR 산물로인한위양성의해결책 Specifications Enzyme: Top DNA polymerase 5 to 3 exonuclease activity: No 3 to 5 exonuclease activity: No 3 - A overhang: Yes Fragment size: ~ 10 kb Application Conventional PCR Primer extension TA cloning Gene sequencing Molecular diagnosis Description AccuPower PCR PreMix (with UDG) 는임상진단이나실험시, PCR 반응에있어심각한문제를야기할수있는 carryover contamination 을최소화할수있는제품입니다. PCR 은민감하고신속한검사법중하나이지만, 이전에증폭된 PCR 산물의오염에의해위양성의문제가흔히발생할수있습니다. 본제품은 Uracil DNA Glycosylase system 을도입하여이러한문제점을극복했습니다. Storage Temperature -20 Features and Benefits Prevention of Carryover Contamination: Uracil DNA Glycosylase (UDG) 는 DNA 가닥내 uracil 과 deoxyribose 간의 N-glycosylic bond 를가수분해함으로써 uracil 이삽입된 template 을제거합니다. UDG 는 PCR cycling 전 37 에서 2 분반응을통해활성화되어, uracil 이들어간 DNA 를제거하므로공기중남아있던산물의유입에의한 carryover 오염을방지할수있으며, PCR cycling 중고온에서불활성화됩니다 (Figure 1). Ease-of-use: Uracil DNA Glycosylase, dutp, DNA polymerase, dntps 를포함하여 PCR 수행에필요한모든반응물이 1 회분량씩동결건조형태의 PreMix 제품으로개별포장되어있습니다. 실험자는 template DNA, primer set, D.W. 만넣어바로 PCR 반응을수행할수있습니다. 또한전기영동시에필요한 tracking dye 와침강제가포함되어있어별도로 sample loading buffer 를첨가할필요가없으므로간편하게사용할수있습니다. Figure 1. Prevention of carryover contamination. Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다 www.bioneer.co.kr 9

AccuPower PCR PreMix (with UDG) Experimental Data PCR PreMix PCR PreMix(with UDG) M 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 PCR PreMix M 1 2 3 4 5 6 7 8 9 10 N PCR PreMix(with UDG) 1 2 3 4 5 678 9 10 N Figure 2. Comparison of sensitivity between AccuPower PCR PreMix and AccuPower PCR PreMix (with UDG). AccuPower PCR PreMix and the competing products (solution type s master mix) are incubated at 95 with various times. Sensitivity test was operated using serial diluted human genomic DNA. Reaction mixture was incubated at 37 for 2 min followed by 95 for 5 min, 30 cycles of 20 sec at 95, 20 sec at 55, 30 sec at 72. Lane 1: 10 ng of human genomic DNA Lane 2: 1 ng of human genomic DNA Lane 3: 100 pg of human genomic DNA Lane 4: 10 pg of human genomic DNA M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030-1) Figure 3. Efficiency of uracil DNA glycosylase using PCR products (including uracil base). Efficiency test of uracil DNA glycosylase was operated using serial diluted PCR products including uracil base. AccuPower PCR PreMix was also tested for negative control. Reaction mixture was incubated at 37 for 2 min, followed by 95 for 5 min, 30 cycles of 20 sec at 95, 20 sec at 55, 30 sec at 72. Lane 1: 10 11 copy Lane 2: 10 10 copy Lane 3: 10 9 copy Lane 4: 10 8 copy Lane 5: 10 7 copy Lane 6: 10 6 copy Lane 7: 10 5 copy Lane 8: 10 4 copy Lane 9: 10 3 copy Lane 10: 10 2 copy Lane N: No template control M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030-1) Ordering Information Cat. no. K-2012-1 K-2016-1 Product Description AccuPower PCR PreMix (with UDG), 0.2 ml thin-wall tubes with attached cap / 96 tubes, 20 μl rxn AccuPower PCR PreMix (with UDG), 0.2 ml thin-wall tubes with attached cap / 480 tubes, 20 μl rxn 10

AccuPower Taq PCR PreMix 동결건조 PCR 제품의 Global Standard Specifications Enzyme: Taq DNA polymerase 5 to 3 exonuclease: Yes 3 to 5 exonuclease: No 3 - A overhang: Yes Fragment Size: ~10kb Application Conventional PCR Primer extension TA cloning Gene sequencing Description AccuPower Taq PCR PreMix 는바이오니아의특허기술로 Taq DNA polymerase, dntps, reaction buffer 등 PCR 수행에필요한구성성분을혼합하여 1 회분량씩동결건조시킨제품입니다. 안정화물질이첨가되어있어상온 (25 ) 보관할경우에는한달동안, 냉동보관할경우에는 2 년동안활성이안정적으로유지되는획기적인제품입니다. Storage Temperature -20 Experimental Data M 1 2 3 4 Supplier I 1 2 3 4 Supplier Q 1 2 3 4 Supplier S 1 2 3 4 Supplier T 1 2 3 4 Features and Benefits Stability: AccuPower Taq PCR PreMix 는 PCR 반응혼합액에안정화제를첨가하여건조시킨제품으로열안전성이떨어지는 solution type 제품의단점을보완하였으며, 장기간보관하더라도활성을안정적으로유지합니다. 또한고온에서도 DNA polymerase 의활성이안정적으로유지되어실험의안정성이확보됩니다. Ease-of-use: 각각의 PCR tube 에 DNA polymerase 와 PCR 반응에필요한모든구성성분이포함되어있어 template DNA, primer set, D.W. 만넣어바로 PCR 을수행할수있습니다. 또한전기영동시에필요한 tracking dye 와침강제가포함되어있어별도로 sample loading buffer 를첨가할필요가없으므로간편하게사용할수있습니다. Figure 1. Comparison of PCR amplification quality between AccuPower Taq PCR PreMix from Bioneer and other suppliers Taq PCR master mix. Target Gene: Human insulin receptor gene. The cycling conditions for AccuPower Taq PCR PreMix were 95 for 5 min, 30 cycles of 20 sec at 95, 20 sec at 55, and 30 sec at 72. PCR reactions using other suppliers PCR master mix were performed according to each supplier s protocol. Lane 1: 10 ng Human genomic DNA Lane 2: 1 ng Human genomic DNA Lane 3: 100 pg Human genomic DNA Lane 4: 10 pg Human genomic DNA Lane M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030) Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. Sensitivity: 뛰어난민감도와증폭효율로미량의 human gdna target 을검출합니다. 타사제품과의비교실험을통해우수한민감도를확인했습니다. www.bioneer.co.kr 11

AccuPower Taq PCR PreMix Supplier I M 1 2 3 4 1 2 3 4 Supplier S 1 2 3 4 Supplier Q 1 2 3 4 Supplier T 1 2 3 4 Figure 2. Comparison of amplification quality between AccuPower Taq PCR PreMix and competitors Taq PCR master mix. Target Gene: IRGC (Immunity-related GTPase family, cinema). Bioneer reaction mixture was followed by 95 for 5 min, 35 cycles of 20 sec at 95, 20 sec at 55, 30 sec at 72. PCR reactions using other suppliers PCR master mix were performed according to each supplier s protocol. Lane 1: 10 ng human genomic DNA Lane 2: 1 ng human genomic DNA Lane 3: 100 pg human genomic DNA Lane 4: 10 pg human genomic DNA Lane M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030) M 1 2 3 4 Supplier I 1 2 3 4 Supplier Q 1 2 3 4 Supplier S 1 2 3 4 Supplier T 1 2 3 4 M Figure 3. Comparison of long kb amplification between AccuPower Taq PCR PreMix and competitors Taq PCR master mix. Bioneer reaction mixture was followed by 95 for 5 min, 30 cycles of 20 sec at 95, 20 sec at 65, 8 min at 68. PCR reactions using other supplier s PCR master mix were performed according to each suppliers protocol. Lane 1: 3 kb fragment (Human tumor protein p53 gene) Lane 2: 4 kb fragment (Human beta globin region) Lane 3: 4.5 kb fragment (Human DNA cross-link repair 1A gene) Lane 4: 8 kb fragment (Human hemoglobin epsilon 1 gene) Lane M: 1 kb DNA Ladder (Bioneer, Cat. no. D-1040) Ordering Information Cat. no. Product Description K-2601 AccuPower Taq PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 20 μl rxn, K-2602 AccuPower Taq PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 20 μl rxn K-2603 AccuPower Taq PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 50 μl rxn K-2604 AccuPower Taq PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 50 μl rxn K-2609 Taq PCR Master Mix, 2.5 ml of 2X master mix solution K-2610 Taq PCR Master Mix, 25 ml of 2X master mix solution, 12.5 ml X 2 tubes 12

AccuPower HotStart PCR PreMix 세계적인바이오니아특허기술을적용한 High Specificity 용 HotStart PCR Kit Specifications Enzyme: Top DNA polymerase 5 to 3 exonuclease activity: No 3 to 5 exonuclease activity: No 3 - A overhang: Yes Fragment size: ~ 12 kb Description AccuPower HotStart PCR PreMix 는바이오니아의세계적인특허기술인 enzyme-mediated HotStart 방법을적용하여반응특이성과 PCR 증폭효율을높인제품입니다. 본제품은 Top DNA polymerase 가낮은온도에서반응하면서발생할수있는 mis-priming, primer-dimer 와같은비특이반응을줄일수있으므로, Complex gdna 또는 cdna templates, Lowcopy targets 과 Multiple primer pairs 등다양한 PCR 증폭반응에사용하실수있습니다. Application High specificity PCR High sensitivity PCR gdna template PCR Low copy target PCR Multiple primer pairs PCR cdna template PCR Storage Temperature -20 Features and Benefits Specificity: 바이오니아의특허기술 (enzyme-mediated HotStart 방법 ) 을적용하여반응전 (zero cycle) 의비특이증폭산물생성을최소화하고, 매 cycle 마다생성되는 PCR inhibitor (PPi) 를가수분해하여 PCR 반응효율을극대화함으로써미량의 template DNA 를효과적으로증폭할수있습니다 (Figure 1). Figure 1. Enzyme-mediated HotStart PCR. Sensitivity: 뛰어난민감도와증폭효율로미량의 human gdna target 을검출합니다. 타사제품과의비교실험을통해우수한민감도를확인했습니다. Ease-of-use: 각각의 PCR tube 에 DNA polymerase 와 PCR 수행에필요한모든구성성분이포함되어있어 template DNA, primer set, D.W. 만넣어바로 PCR 반응을수행할수있습니다. 또한전기영동시에필요한 tracking dye 와침강제가포함되어있어 sample loading buffer 를첨가할필요가없으므로간편하게사용할수있습니다. Stability: PCR 반응혼합물에안정화제를첨가하여건조시켜, 장기간보관하더라도활성을안정적으로유지합니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. www.bioneer.co.kr 13

AccuPower HotStart PCR PreMix Experimental Data Figure 2. Specificity comparison between AccuPower PCR PreMix and AccuPower HotStart PCR PreMix. Lane 1: P75/P73 primer set (139 bp) Lane 2: P55/P53 primer set (211 bp) Lane 3: P55/P63 primer set (447 bp) Lane 4: P75/P83 primer set (618 bp) Lane 5: P55/P73 primer ser (1082 bp) Lane 6: P65/P83 primer set (1296 bp) Lane 7: P55/P83 primer set (1561 bp) Figure 3. Comparison of PCR amplification sensitivity between Accu- Power HotStart PCR PreMix from Bioneer and other suppliers Hot- Start PCR kit. Lane 1: Human gdna 10 ng Lane 2: Human gdna 1 ng Lane 3: Human gdna 100 pg Lane 4: Human gdna 10 pg M: 100 bp plus DNA ladder (Bioneer, Cat. no D-1035) Ordering Information Cat. no. Product Description K-5050 AccuPower HotStart PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 20 μl rxn, K-5051 AccuPower HotStart PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 20 μl rxn K-5052 AccuPower HotStart PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 50 μl rxn K-5053 AccuPower HotStart PCR PreMix, 0.5 ml thin-wall tubes with attached cap / 100 tubes, 50 μl rxn K-5057 AccuPower HotStart PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 50 μl rxn 14

AccuPower HotStart PCR PreMix (with UDG) 비특이적반응과 Carryover Contamination 을제거하는데최적화된 Kit Specifications Enzyme: Top DNA polymerase 5 to 3 exonuclease activity: No 3 to 5 exonuclease activity: No 3 - A overhang: Yes Fragment size: ~ 12 kb Application gdna template PCR Low-copy target PCR Multiple primer pairs PCR cdna template PCR Molecular diagnosis Description AccuPower HotStart PCR PCR PreMix (with UDG) 는 enzymemediated HotStart 기술적용으로 Primer-dimer 형성을최소화하여반응특이성과 PCR 증폭효율을높임과동시에 uracil DNA glycosylase system 적용으로 carryover contamination 문제를해결한제품입니다. Storage Temperature -20 Features and Benefits Prevention of Carryover Contamination: Uracil DNA Glycosylase (UDG) 는 DNA 가닥내 uracil과 deoxyribose 간의 N-glycosylic bond를가수분해함으로써 uracil이삽입된 template을제거합니다. UDG는 PCR cycling 전 37 에서 2분반응을통해활성화되어, uracil이들어간 DNA를제거하므로공기중남아있던산물의유입에의한 carryover 오염을방지할수있으며, PCR cycling 중고온에서불활성화됩니다 (Figure 1). Specificity: PPi 와 PPase 가포함된 enzyme-mediated HotStart 제품은 PCR 반응의부산물인 PPi 를 Pi 로분해함으로써반응을증가시키는효과를가지고있어서비특이적증폭산물생성을억제하고, 특이적증폭산물을효과적으로생성하여미량의 target DNA 에대한증폭반응, 여러서열 primer 를사용한증폭반응, PCR 반응시발생하는비특이적증폭산물억제에매우유용하게사용할수있습니다. Figure 1. Prevention of carryover contamination. Ease-of-use: 각각의 PCR tube 에 DNA polymerase 와 PCR 수행에필요한모든구성성분이포함되어있어 template DNA, primer set, D.W. 만넣어바로 PCR 반응을수행할수있습니다. 또한전기영동시에필요한 tracking dye 와침강제가포함되어있어 sample loading buffer 를추가로넣을필요가없으므로간편하게사용할수있습니다. Stability: PCR 반응혼합물에안정화제를첨가하여건조시켜, 장기간보관하더라도활성을안정적으로유지합니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거쳐재현성있는실험결과를제공합니다. www.bioneer.co.kr 15

AccuPower HotStart PCR PreMix (with UDG) Experimental Data Figure 2. Comparison of specificity between AccuPower HotStart PCR PreMix and HotStart PCR PreMix (with UDG). Specificity test was operated using 7 pairs of primers targeting P53 gene. Reaction mixture was incubated at 37 for 2 min followed by 95 for 5 min, 30 cycles of 20 sec at 95, 40 sec at 55, 1 min at 72. The amount of DNA (human) used to test is 10 ng. Lane 1: 139 bp Lane 2: 211 bp Lane 3: 447 bp Lane 4: 618 bp Lane 5: 1082 bp Lane 6: 1,296 bp Lane 7: 1,561 bp M: 100 bp plus DNA Ladder (Bioneer, Cat. no. D-1030-1) Figure 3. Efficiency of uracil DNA glycosylase using PCR product (including uracil base). Efficiency test of uracil DNA glycosylase was operated using serial diluted PCR products including uracil base. AccuPower HotStart PCR PreMix was also tested for negative control. Reaction mixture was incubated at 37 for 2 min followed by 95 for 5 min, 30 cycles of 20 sec at 95, 20 sec at 55, 30 sec at 72. Lane 1: 10 11 copy Lane 2: 10 10 copy Lane 3: 10 9 copy Lane 4: 10 8 copy Lane 5: 10 7 copy Lane 6: 10 6 copy Lane 7: 10 5 copy Lane 8: 10 4 copy Lane N: No template control M: 100 bp plus DNA Ladder (Bioneer, Cat. no. D-1030-1) Ordering Information Cat. no. Product Description K-5050-1 AccuPower HotStart PCR PreMix (with UDG), 0.2 ml thin-wall tubes with attached cap / 96 tubes, 20 μl rxn K-5051-1 AccuPower HotStart PCR PreMix (with UDG), 0.2 ml thin-wall tubes with attached cap / 480 tubes, 20 μl rxn 16

AccuPower GoldHotStart Taq PCR PreMix NEW HotStart 를적용한 High Specificity Kit Specifications Enzyme: GoldHotStart Taq DNA polymerase 5 to 3 exonuclease: Yes 3 to 5 exonuclease: No 3 - A overhang: Yes Fragment size: ~ 5 kb (Human) Description AccuPower GoldHotStart Taq PCR Premix 는 GoldHotStart Taq DNA polymerase 를적용하여 hybridization efficiency 를향상시킴으로써반응특이성과 PCR 증폭효율을높인제품입니다. 본제품은 Taq DNA polymerase 가낮은온도에서반응하면서발생할수있는 nonspecific-priming, primer-dimer 와같은비특이반응을줄여줍니다. Features and Benefits Specificity: GoldHotStart Taq DNA polymerase 는반응전 (zero cycle) 의비특이증폭산물의생성을최소화합니다. PCR 온도가증가하면서활성화된 GoldHotStart Taq DNA polymerase 는정확한 target 만을빠르게증폭합니다. Sensitivity: 뛰어난민감도와증폭효율로미량의 human gdna target 을검출합니다. 타사제품과의비교실험을통해우수한민감도를확인했습니다. Ease-of-use: 각각의 PCR tube 에 DNA polymerase 와 PCR 수행에필요한모든구성성분이포함되어있어 template DNA, primer set, D.W. 만넣어바로 PCR 반응을수행할수있습니다. 또한전기영동시에필요한 tracking dye 와침강제가포함되어있어 sample loading buffer 를추가로넣을필요가없으므로간편하게사용할수있습니다. Stability: PCR 반응혼합물에안정화제를첨가하여건조시켜, 장기간보관하더라도활성을안정적으로유지합니다. Application High specificity PCR High sensitivity PCR gdna template PCR Low-copy target PCR Multiple primer pairs PCR cdna template PCR TA cloning Storage Temperature -20 Experimental Data Figure 1. Comparison of PCR amplification efficiency between Accu- Power GoldHotStart Taq PCR PreMix from Bioneer and other suppliers HotStart PCR master mix. Target: Human insulin receptor gene. The cycling conditions for AccuPower GoldHotStart Taq PCR PreMix were 95 for 5 min, 30 cycles of 95 for 30 sec, 55 for 30 sec and 72 for 30 sec. PCR reactions using other suppliers PCR master mix were performed according to each suppliers protocol. Lane 1: 10 ng of human gdna Lane 2: 1 ng of human gdna Lane 3: 100 pg of human gdna Lane 4: 10 pg of human gdna Lane M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030) Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. www.bioneer.co.kr 17

AccuPower GoldHotStart Taq PCR PreMix NEW Experimental Data Figure 2. Comparison of PCR amplification specificity between AccuPower GoldHotStart Taq PCR PreMix from Bioneer and other suppliers Hotstart PCR master mix. Target: ApoE gene (The PCR product size is 268 bp). The cycling conditions for AccuPower GoldHotStart Taq PCR PreMix were 95 for 5 min, 35 cycles of 95 for 30 sec, 57 for 30 sec and 72 for 30 sec. PCR reactions using other suppliers PCR master mix were performed according to each supplier s protocol. Lane 1,2: 100 ng human genomic DNA Lane 3,4: 10 ng human genomic DNA Lane M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030) Figure 3. Comparison of PCR amplification specificity between AccuPower GoldHotStart Taq PCR PreMix from Bioneer and other suppliers Hotstart PCR master mix. The cycling conditions for AccuPower GoldHotStart Taq PCR PreMix were 95 for 5 min, 35 cycles of 95 for 30 sec, 65 for 30 sec and 72 for 30 sec. PCR reactions using other suppliers PCR master mix were performed according to each supplier s protocol. Lane 1: 100 ng human genomic DNA Lane 2: 10 ng human genomic DNA Lane M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030) Ordering Information Cat. no. Product Description K-2621 AccuPower GoldHotStart Taq PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 20 µl rxn K-2622 AccuPower GoldHotStart Taq PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 20 µl rxn K-2623 AccuPower GoldHotStart Taq PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 50 µl rxn K-2624 AccuPower GoldHotStart Taq PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 50 µl rxn K-2629 AccuPower GoldHotStart Taq PCR MasterMix, 2.5 ml of 2X master mix solution K-2630 AccuPower GoldHotStart Taq PCR MasterMix, 25 ml of 2X master mix solution 18

AccuPower PyroHotStart Taq PCR PreMix 세계적인바이오니아특허기술을적용한 High Specificity 용 Kit Specifications Enzyme: Taq DNA polymerase 5 to 3 exonuclease: Yes 3 to 5 exonuclease: No 3 - A overhang: Yes Fragment size: ~ 5 kb (Human) Description AccuPower PyroHotStart Taq PCR PreMix 는바이오니아의세계적인특허기술인 enzyme-mediated HotStart 방법을적용하여반응특이성과 PCR 증폭효율을높인제품입니다. 본제품은 Taq DNA polymerase 가낮은온도에서반응하면서발생할수있는 mis-priming, primer-dimer 와같은비특이반응을줄여줍니다. Application High specificity PCR High sensitivity PCR gdna template PCR Low-copy target PCR Multiple primer pairs PCR cdna template PCR TA cloning Storage Temperature -20 Features and Benefits Specificity: 바이오니아의특허기술 (enzyme-mediated HotStart 방법 ) 을적용하여반응전 (zero cycle) 의비특이증폭산물생성을최소화하고, 매 cycle 마다생성되는 PCR inhibitor (PPi) 를가수분해하여 PCR 반응효율을극대화함으로써미량의 template DNA 를효과적으로증폭할수있습니다 (Figure 1). Sensitivity: 뛰어난민감도와증폭효율로미량의 human gdna target 을검출합니다. 타사제품과의비교실험을통해우수한민감도를확인했습니다. Ease-of-use: 각각의 PCR tube 에 DNA polymerase 와 PCR 수행에필요한모든구성성분이포함되어있어 template DNA, primer set, D.W. 만넣어바로 PCR 반응을수행할수있습니다. 또한전기영동시에필요한 tracking dye 와침강제가포함되어있어 sample loading buffer 를추가로넣을필요가없으므로간편하게사용할수있습니다. Figure 1. Enzyme-mediated HotStart PCR. Stability: PCR 반응혼합물에안정화제를첨가하여건조시켜, 장기간보관하더라도활성을안정적으로유지합니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. www.bioneer.co.kr 19

AccuPower PyroHotStart Taq PCR PreMix Experimental Data Supplier I M 1 2 3 4 1 2 3 4 Supplier S 1 2 3 4 Supplier T 1 2 3 4 Supplier Q 1 2 3 4 Figure 2. Comparison of PCR amplification specificity between AccuPower PyroHotStart Taq PCR PreMix from Bioneer and other suppliers hotstart PCR master mix PCR reactions were performed according to each supplier s protocol. Target: Human Prp gene The PrP gene was amplified from human genomic DNA with two different primer sets, separately. This data shows that AccuPower PyroHotStart Taq PCR PreMix has higher amplificition efficency and specificity than other suppliers HotStart PCR master mix. Lane 1: 100 ng DNA PrP primer set (500 bp) Lane 2: 10 ng DNA PrP primer set (500 bp) Lane 3: 100 ng DNA PrP primer set (705 bp) Lane 4: 10 ng DNA PrP primer set (705 bp) Lane M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030) Figure 4. AccuPower PyroHotStart Taq PCR PreMix has high amplification efficiency and specificity. Specificity test was performed using 7 different sets of primers targeting the P53 gene. 10 ng of human genomic DNA was used for each PCR reaction. The cycling conditions were 95 for 5 min, 30 cycles of 95 for 20 sec, 55 for 40 sec, and 72 for 1 min, and 72 for 5 min for final extension. Lane 1: P75/73 primer set (139 bp) Lane 2: P55/53 primer set (211 bp) Lane 3: P55/63 primer set (447 bp) Lane 4: P75/83 primer set (618 bp) Lane 5: P55/73 primer set (1,082 bp) Lane 6: P65/83 primer set (1,296 bp) Lane 7: P55/83 primer set (1,561 bp) Lane M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030) Figure 3. Comparison of PCR amplification specificity between AccuPower PyroHotStart Taq PCR PreMix from Bioneer and other suppliers HotStart PCR master mix. The ApoE gene was amplifed from 100 ng of human genomic DNA (The PCR product size is 268 bp). This data shows that AccuPower PyroHotStart Taq PCR PreMix has higher amplificition efficency and specificity than other suppliers HotStart PCR master mix. Lane 1: AccuPower PyroHotStart Taq PCR PreMix Lane 2: Supplier I HotStart Taq PCR premix Lane 3: Supplier S HotStart Taq PCR master mix Lane 4: Supplier T HotStart Taq PCR master mix Lane 5: Supplier Q HotStart Taq PCR master mix Lane M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030) Ordering Information Cat. no. Product Description K-2611 AccuPower PyroHotStart Taq PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 20 μl rxn K-2612 AccuPower PyroHotStart Taq PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 20 μl rxn K-2613 AccuPower PyroHotStart Taq PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 50 μl rxn K-2614 AccuPower PyroHotStart Taq PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 50 μl rxn 20

AccuPower HotStart Pfu PCR PreMix Gene Cloning 등 High Fidelity 를요구하는실험에최적화된 Kit Specifications Enzyme: Pfu DNA polymerase 5 to 3 exonuclease activity: No 3 to 5 exonuclease activity: Yes 3 - A overhang: No Fragment size: ~ 10 kb Application Gene cloning with blunt ends Site-directed mutagenesis High fidelity amplification High specificity PCR cdna template PCR Description AccuPower HotStart Pfu PCR PreMix 는세계적으로인정받은바이오니아특허기술인 enzyme-mediated HotStart 기술이적용되어 miss-priming, primer dimer, non-specific amplification 을근본적으로제거할수있어서미량의 target DNA 도정확하게증폭해낼수있는제품입니다. 또한본제품에포함된 Pfu DNA polymerase 는 proof-readiing (3 5 exonuclease) 기능을가지고있어서 DNA 증폭반응시발생하는 error 를줄일수있어단백질생산을위한 cloning 실험에적합합니다. Storage Temperature -20 Experimental Data Features and Benefits High Fidelity: Proof-reading 기능과높은특이성을가지고있어 cloning 에가장적합한제품입니다. Ease-of-use: 각각의 PCR tube 에 DNA polymerase 와 PCR 수행에필요한모든구성성분이포함되어있어 template DNA, primer set, D.W. 만넣어바로 PCR 반응을수행할수있습니다. 또한전기영동시에필요한 tracking dye 와침강제가포함되어있어 sample loading buffer 를추가로넣을필요가없으므로간편하게사용할수있습니다. Specificity: Pfu DNA polymerase 의 proof-reading 기능을최대한살리면서 enzyme-mediated HotStart 의특장점을통해 cloning, mutagenesis 등정확한 sequence 가요구되는실험에서최적의 PCR product 를얻을수있습니다. Stability: PCR 반응혼합물에안정화제를첨가하여건조시켜, 장기간보관하더라도활성을안정적으로유지합니다. Figure 1. AccuPower HotStart Pfu PCR PreMix shows enhanced specificity compared to competitors. Specificity test was performed using 7 different sets of primers targeting the p53 gene. 10 ng of human genomic DNA was used for each PCR reaction. The cycling conditions were 95 for 5 min, 32 cycles of 95 for 30 sec, 62 for 40 sec, and 72 for 1 min 30 sec, and 72 for 5 min for final extension. Lane 1: P75/73 primer set (139 bp) Lane 2: P55/53 primer set (211 bp) Lane 3: P55/63 primer set (447 bp) Lane 4: P75/83 primer set (618 bp) Lane 5: P55/73 primer set (1082 bp) Lane 6: P65/83 primer set (1296 bp) Lane 7: P55/83 primer set (1561 bp) Lane M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030) Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. www.bioneer.co.kr 21

AccuPower HotStart Pfu PCR PreMix Figure 2. AccuPower HotStart Pfu PCR PreMix has high amplification efficiency. Bioneer reaction mixture was followed by 95 for 5 min, 35 cycles of 95 for 20 sec, 65 for 20 sec, and 68 for 15 min, and 68 for 5 min for final extension. Lane 1: 2 kb fragment Lane 2: 2.5 kb fragment Lane 3: 3 kb fragment Lane 4: 4.5 kb fragment Lane 5: 5 kb fragment Lane M: 1 kb DNA Ladder (Bioneer, Cat. no. D-1040) Ordering Information Figure 3. Comparison of PCR amplification efficiency between AccuPower HotStart Pfu PCR PreMix from Bioneer and other suppliers PCR master mix Target: human insulin receptor gene. The cycling conditions for AccuPower HotStart Pfu PCR PreMix were 95 for 5 min, 30 cycles of 95 for 30 sec, 55 for 30 sec and 72 for 2 min. PCR reactions using other suppliers PCR master mix were performed according to each supplier s protocol. Lane 1: 10 ng of human genomic DNA Lane 2: 1 ng of human genomic DNA Lane 3: 100 pg of human genomic DNA Lane 4: 10 pg of human genomic DNA Lane M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030) Cat. no. Product Description K-2301 AccuPower HotStart Pfu PCR PreMix 96 tubes, 20 μl rxn K-2302 AccuPower HotStart Pfu PCR PreMix 96 tubes, 50 μl rxn K-2303 AccuPower HotStart Pfu PCR PreMix 480 tubes, 20 μl rxn K-2304 AccuPower HotStart Pfu PCR PreMix 480 tubes, 50 μl rxn 22

AccuPower Pfu PCR PreMix Gene Cloning 등 High Fidelity 를요구하는실험에최적화된 Kit Description AccuPower Pfu PCR PreMix 는 proof-reading 기능을갖는 Pfu DNA polymerase 를사용하여매우정확한 PCR product 을얻을수있는제품입니다. 또한, Pfu DNA polymerase 와 PCR 반응혼합물이동결건조되어제공되므로 template DNA 와 primers 의첨가만으로간편하게 high fidelity 를갖는 PCR product 를얻을수있습니다. Specifications Enzyme: Pfu DNA polymerase 5 to 3 exonuclease activity: No 3 to 5 exonuclease activity: Yes 3 - A overhang: No Fragment size: ~ 15 kb Application Gene synthesis Gene cloning Conventional PCR Primer extension Site directed mutagenesis High fidelity 가요구되는실험 Storage Temperature -20 Features and Benefits High Fidelity: 높은정확성 (error rate= 1.9X10-6 ) 을가지고있어 DNA 증폭시발생하는 mutation 을최소화하였습니다. Sensitivity: 높은민감도와증폭효율로미량의 human gdna target 을검출합니다. Long Range PCR: Human gdna 의경우 15 kb 이상의 PCR 를효과적으로증폭시킬수있습니다. Ease-of-use: 각각의 PCR tube 에 DNA polymerase 와 PCR 수행에필요한모든구성성분이포함되어있어 template DNA, primer set, D.W. 만넣어바로 PCR 반응을수행할수있습니다. 또한전기영동시에필요한 tracking dye 와침강제가포함되어있어 sample loading buffer 를첨가할필요가없으므로간편하게사용할수있습니다. Stability: PCR 반응혼합물에안정화제를첨가하여건조시켜, 장기간보관하더라도활성을안정적으로유지합니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. www.bioneer.co.kr 23

AccuPower Pfu PCR PreMix Experimental Data Figure 1. Template range & sensitivity of AccuPower Pfu PCR PreMix for human DNA template. Test of working range & sensitivity of AccuPower Pfu PCR PreMix for human DNA template. Line 1: 100 ng Line 2: 10 ng Line 3: 1 ng Line 4: 100 pg Line 5: 10 pg Line 6: Template negative M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030) Figure 2. Amplification of lambda DNA of 1 kb to 10 kb with AccuPower Pfu PCR PreMix. Lane 1: 1 kb fragment Lane 2: 2 kb fragment Lane 3: 3 kb fragment Lane 4: 4 kb fragment Lane 5: 5 kb fragment Lane 6: 6 kb fragment Lane 7: 7 kb fragment Lane 8: 8 kb fragment Lane 9: 9 kb fragment Lane 10: 10 kb fragment M: 1 kb DNA Ladder (Bioneer, Cat. no. D-1040) Ordering Information Cat. no. Product Description K-2022 AccuPower Pfu PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 20 μl rxn K-2023 AccuPower Pfu PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 50 μl rxn K-2024 AccuPower Pfu PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 20 μl rxn K-2025 AccuPower Pfu PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 50 μl rxn K-2026 AccuPower Pfu PCR Master Mix, 1 ml of 2X master mix solution K-2027 AccuPower Pfu PCR PreMix, 0.5 ml thin-wall tubes with attached cap / 100 tubes, 50 μl rxn 24

AccuPower ProFi Taq PCR PreMix High Efficiency 및 Long Range PCR 에최적화된 Kit Specifications Enzyme: ProFi Taq DNA polymerase 5 to 3 exonuclease: Yes 3 to 5 exonuclease: Yes 3 - A overhang: Yes PCR product size: ~ 30 kb Description AccuPower ProFi Taq PCR PreMix 는바이오니아에서독자적으로개발한 ProFi Taq DNA polymerase 을사용하여높은정확성과증폭효율로긴 template DNA 증폭에최적화된제품입니다. 또한, 본제품은 ProFi Taq DNA polymerase 와 PCR 반응혼합물이동결건조되어제공되므로 template DNA 와 primers 의첨가만으로정확하고깨끗한 PCR 산물을얻을수있습니다. Features and Benefits Long Range PCR: Lambda DNA 의경우 30 kb, human gdna 의경우 21 kb 의 PCR 산물을효과적으로증폭할수있습니다. Sensitivity: 뛰어난민감도와증폭효율로미량의 human gdna target 을검출합니다. 타사제품과의비교실험을통해우수한민감도를확인했습니다. Ease-of-use: 각각의 PCR tube 에 ProFi Taq DNA polymerase 와 PCR 반응에필요한모든구성성분이포함되어있어 template DNA, primer set 와 D.W. 만넣어바로 PCR 반응을수행할수있습니다. 또한전기영동시에필요한 tracking dye 와침강제가포함되어있어별도로 sample loading buffer 를첨가할필요가없으므로간편하게사용할수있습니다. Stability: PCR 반응혼합물에안정화제를첨가하여건조시켜, 장기간보관하더라도활성을안정적으로유지합니다. Application Primer extension Long range amplification from genomic DNA High amplification efficiency Excellent performance on difficult templates Amplification of low-copy targets High yield and high sensitivity PCR Storage Temperature -20 Experimental Data M 1 2 3 4 Supplier I 1 2 3 4 Supplier S 1 2 3 4 Supplier T 1 2 3 4 Figure 1. Comparison of PCR amplification efficiency between AccuPower ProFi Taq PCR PreMix from Bioneer and other suppliers PCR master mix. cdna synthesized from 10-fold serial-diluted human total RNA from 10 ng to 10 pg using AccuPower RocketScript TM Cycle RT PreMix(Bioneer, Cat. no. K-2201) was used as a template for PCR amplification. The cycling conditions for AccuPower ProFi Taq PCR PreMix were 95 for 5 min, 33 cycles of 95 for 20 sec, 55 for 20 sec and 72 for 30 sec. PCR reactions using other suppliers PCR master mix were performed according to each supplier s protocol. Target: human GAPDH gene Lane 1: 10 ng of human total cdna Lane 2: 1 ng of human total cdna Lane 3: 100 pg of human total cdna Lane 4: 10 pg of human total cdna Lane M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030) Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. www.bioneer.co.kr 25

AccuPower ProFi Taq PCR PreMix M 1 2 3 4 Supplier I 1 2 3 4 Supplier S 1 2 3 4 Supplier T 1 2 3 4 M M1M2 1 2 3 4 Supplier I 1 2 3 4 Supplier S 1 2 3 4 Supplier T 1 2 3 4 M2M1 Figure 2. Comparison of PCR amplification sensitivity between AccuPower ProFi Taq PCR PreMix from Bioneer and other suppliers PCR master mix. The cycling conditions for AccuPower ProFi Taq PCR PreMix were 95 for 5 min, 30 cycles of 95 for 20 sec, 65 for 20 sec and 68 for 4 min. PCR reactions using other suppliers PCR master mix were performed according to each supplier s protocol. Lane 1: 2 kb fragment (human tumor protein p53 gene) Lane 2: 3 kb fragment (human tumor protein p53 gene) Lane 3: 4.5kb fragment (human DNA cross-link repair 1A gene) Lane 4: 8 kb fragment (human hemoglobin epsilon 1 gene) Lane M: 1 kb DNA Ladder (Bioneer, Cat. no. D-1040) Figure 3. Comparison of PCR amplification of long targets between AccuPower ProFi Taq PCR PreMix from Bioneer and other suppliers PCR master mix. The cycling conditions for AccuPower ProFi Taq PCR PreMix were 95 for 5 min, 32 cycles of 95 for 20 sec, 65 for 40 sec, and 68 for 15 min. PCR reactions using other suppliers PCR master mix were performed according to each supplier s protocol. Human DNA was used as a template for PCR amplification. Lane 1: 11 kb fragment Lane 2: 13.5 kb fragment Lane 3: 17.6 kb fragment Lane 4: 21.4 kb fragment Lane M1: Lambda/Hind III marker (Bioneer, Cat. no. D-1050) Lane M2: 1 kb DNA Ladder (Bioneer, Cat. no. D-1040) Ordering Information Cat. no. Product Description K-2631 AccuPower ProFi Taq PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached caps / 96 tubes, 20 μl rxn/tube K-2632 AccuPower ProFi Taq PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached caps / 480 tubes, 20 μl rxn/tube K-2633 AccuPower ProFi Taq PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached caps / 96 tubes, 50 μl rxn/tube K-2634 AccuPower ProFi Taq PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached caps / 480 tubes, 50 μl rxn/tube 26

AccuPower Multiplex PCR PreMix 하나의 Tube 내에서 Multiplex PCR (Target gene 20 종까지 ) 이가능한 Kit Specifications Enzyme: HotStart Top DNA polymerase 5 to 3 exonuclease activity: No 3 to 5 exonuclease activity: No 3 - A overhang: Yes Fragment size: ~ 1 kb Application 유전자진단관련 assay RADP DNA and RNA chip cdna library Description AccuPower Multiplex PCR PreMix 는하나의 tube 내에서한번반응으로 20 개 target gene 의 PCR 산물을얻을수있는획기적인제품입니다. 또한 antibody-based HotStart Top DNA polymerase 를적용하여반응특이성과 PCR 증폭효율을높였습니다. 본제품은 multiplex PCR 기술을이용한일반적인 assay 및유전자진단관련 genotyping assay 에도적용이가능하며, cdna 를이용한 semi-quantitative gene expression 실험에적합합니다. Storage Temperature -20 Experimental Data Features and Benefits Multiplex PCR: Template DNA 를이용하여한 tube 내에서 20 개의 multiplex PCR 증폭산물생성이가능하여경제적이고편리합니다. Specificity: HotStart 가적용된 DNA polymerase 는반응전 (zero cycle) 의비특이증폭산물의생성을최소화합니다. PCR 온도가증가하면서활성화된 DNA polymerase 는정확한 target 만을빠르게증폭합니다. Ease-of-use: 각각의 PCR tube 에 HotStart Top DNA polymerase 와 PCR 수행에필요한모든구성성분이포함되어있어 template DNA, primer set, D.W. 만넣어바로 multiplex PCR 반응을수행할수있습니다. 또한전기영동시에필요한 tracking dye 와침강제가포함되어있어 sample loading buffer 를추가로넣을필요가없으므로편리하게사용할수있습니다. Figure 1. Single PCR and multiplex PCR using AccuPower Multiplex PCR PreMix. Each lane from left to right indicates the single and multiplex PCR product using AccuPower Multiplex PCR PreMix. a) 10-plex multiplex PCR b) 20-plex multiplex PCR M: 25/100 bp Mixed DNA Ladder (Bioneer, Cat. no. D-1020) Stability: PCR 반응혼합물에안정화제를첨가하여건조시켜, 장기간보관하더라도활성을안정적으로유지합니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. www.bioneer.co.kr 27

AccuPower Multiplex PCR PreMix Figure 2. Comparison of amplification quality between AccuPower Multiplex PCR PreMix and other suppliers multiplex PCR kit. 6-plex primers were added into AccuPower Multiplex PCR PreMix and other suppliers multiplex PCR kit. A series of human genomic DNA diluents were tested. (Lane 1: Human genomic DNA 100 ng, Lane 2: Human genomic DNA 10 ng, Lane 3: Human genomic DNA 1 ng). All data were obtained using MyGenie TM 96 Gradient Thermal Block (Bioneer, Cat. no. A-2040-1). Supplier Q: Q company multiplex PCR kit Supplier S: S company multiplex PCR kit Supplier I: I company Taq DNA polymerase for multiplex PCR (0.5 U, added 2 mm MgCl2) Rxn. condition: 95 for 10 min, followed by 35 cycles of 30 sec at 95, 30 sec at 60, 60 sec at 72 M: 25/100 bp Mixed DNA Ladder (Bioneer, Cat. no. D-1020) Figure 3. Comparison of amplification quality between AccuPower Multiplex PCR PreMix and other suppliers multiplex PCR kit. 10-plex primers were added into AccuPower Multiplex PCR PreMix and other suppliers multiplex PCR kit. A series of human genomic DNA diluents were tested. (Lane 1: Human genomic DNA 100 ng, Lane 2: Human genomic DNA 10 ng, Lane 3: Human genomic DNA 1 ng). All data were obtained using MyGenie TM 96 Gradient Thermal Block (Bioneer, Cat. No. A-2040-1). Supplier Q: Q company multiplex PCR kit Supplier S: S company multiplex PCR kit Supplier I: I company Taq DNA polymerase for multiplex PCR (0.5 U, added 2 mm MgCl2) Rxn. condition: 95 for 10 min, followed by 35 cycles of 30 sec at 95, 30 sec at 65, 60 sec at 72 M: 25/100 bp Mixed DNA Ladder (Bioneer, Cat. no. D-1020) Figure 4. Comparison of amplification quality using Labchip TM between AccuPower Multiplex PCR PreMix and other suppliers multiplex PCR kit. a) Virtual Gel Image. Gel image illustrates data reproducibility of the LabChip TM 90 system. b) Overlay of expression level using Bioneer s Multiplex PCR PreMix and other company s multiplex PCR kit. The electropherogram displays the data between 10 PCR products yield using 10-plex primer sets to illustrate the amplification efficiency. c) The graph shows the total concentration of PCR products between AccuPower Multiplex PCR PreMix and other suppliers multiplex PCR kit. Ordering Information Cat. no. Product Description K-2111 AccuPower Multiplex PCR PreMix, 0.2 ml thin-wall tubes with attached cap / 96 tubes, 20 μl rxn K-2112 AccuPower Multiplex PCR PreMix, 0.2 ml thin-wall tubes with attached cap / 96 tubes, 50 μl rxn K-2113 AccuPower Multiplex PCR PreMix, 0.2 ml thin-wall tubes with attached cap / 480 tubes, 20 μl rxn K-2114 AccuPower Multiplex PCR PreMix, 0.2 ml thin-wall tubes with attached cap / 480 tubes, 50 μl rxn 28

AccuPower Gold Multiplex PCR PreMix 한 Tube 에서 20 종의 Target Gene 을증폭시킬수있는 Kit Specifications Enzyme: Top DNA polymerase 5 to 3 exonuclease activity: No 3 to 5 exonuclease activity: No 3 - A overhang: Yes Fragment size: ~ 1 kb Application Description AccuPower Gold Multiplex PCR PreMix 는한 tube 내에서한번반응으로 20 개의 target gene 의 PCR 산물을얻을수있는획기적인제품입니다. 세계적으로인정받은바이오니아독창적특허기술인 enzyme-mediated HotStart 방법을적용하여정확하고깨끗한 multiplex PCR 산물을얻을수있습니다. 본제품은 multiplex PCR 기술을이용한 genotyping assay 나유전자진단등다양한분야에적용이가능합니다. Features and Benefits Multiplex PCR: Template DNA 를이용하여한 tube 내에서 20 개의 multiplex PCR 증폭산물생성이가능하여경제적이고편리합니다. Specificity: PPi 와 PPase 를이용하는 enzyme-mediated HotStart PCR 은반응전 (zero cycle) 의비특이증폭산물의생성을최소화합니다. 반응온도가 70 이상으로올라가면 PPase 가 PPi 를 2 Phosphate (2Pi) 로가수분해하여 Mg 2+ 이온이해리되고, DNA 중합효소는해리된 Mg 2+ 이온을이용하여정확한 target 만을빠르게증폭합니다 (Figure 1). Ease-of-use: 각각의 PCR tube 에 Top DNA polymerase 와 PCR 수행에필요한모든구성성분이포함되어있어 template DNA, primer set, D.W. 만넣어바로 multiplex PCR 반응을수행할수있습니다. 또한전기영동시에필요한 tracking dye 와침강제가포함되어있어 sample loading buffer 를추가로넣을필요가없으므로간편하게사용할수있습니다. Target Human and Animal Plant Storage Temperature -20 Application STR analysis for determining genetic profiles in forensic cases Molecular diagnostic analysis Genotyping assay Qualitative and semi-qualitative gene expression assay Mutant screening Transgenic organism analysis STR analysis Detection of pathogens/bacteria infection Transgenic organism analysis Qualitative and semi-qualitative gene expression assay Figure 1. Enzyme-mediated HotStart PCR. Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. Stability: PCR 반응혼합물에안정화제를첨가하여건조시켜, 장기간보관하더라도활성을안정적으로유지합니다. www.bioneer.co.kr 29

AccuPower Gold Multiplex PCR PreMix Experimental Data Figure 2. High specificity of AccuPower Gold Multiplex PCR PreMix. Each lane from left to right represents the progressive number of primer sets (1~20) included in AccuPower Gold Multiplex PCR PreMix reactions. Rxn. condition: 95 for 10 min, followed by 30 cycles (a), 35 cycles (b) of 30 sec at 95, 30 sec at 57, 60 sec at 72 Figure 3. High specificity of AccuPower Gold Multiplex PCR PreMix. Each lane from left to right indicates the single and multiplex PCR product using AccuPower Gold Multiplex PCR PreMix. a) 10-plex multiplex PCR b) 20-plex multiplex PCR Lane M: 25/100 bp Mixed DNA Ladder (Bioneer, Cat. no. D-1020) Rxn. condition: 95 for 10 min, followed by 30 cycles(a), 35 cycles(b) of 30 sec at 95, 30 sec at 57 C, 60 sec at 72 Figure 4. Comparison of amplification quality between AccuPower Gold Multiplex PCR PreMix and other suppliers multiplex PCR kit. 6-plex a), 10-plex b), 20-plex c) primers were added into AccuPower Gold Multiplex PCR PreMix and other suppliers master mixture. A series of human genomic DNA diluents were tested. (Lane 1: 100 ng, Lane 2: 10 ng, Lane 3: 1 ng). All data were obtained using MyGenie TM 96 Gradient Thermal Block (Bioneer, Cat. no. A-2040-1). Supplier Q: Multiplex PCR master mix Supplier S: Multiplex PCR master mix Supplier I: Taq DNA polymerase for multiplex PCR (0.5 U), added 2 mm MgCl2 M; 25/100 bp Mixed DNA Ladder (Bioneer, Cat. no. D-1020) Rxn. condition: 95 for 10 min, followed by 35 cycles of 30 sec at 95, 30 sec at 60, 60 sec at 72 Ordering Information Cat. no. Product Description K-2115 AccuPower Gold Multiplex PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 20 µl rxn K-2116 AccuPower Gold Multiplex PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 20 µl rxn K-2117 AccuPower Gold Multiplex PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 50 µl rxn K-2118 AccuPower Gold Multiplex PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 50 µl rxn 30

Top DNA Polymerase Enzymes for Everyday PCR, Faster than Taq DNA Polymerase, and TA Cloning Compatible Application Real-Time quantification of DNA and cdna targets using SYBR Green dye. Gene expression profiling Microbial & viral pathogen detection Reagents Supplied 10 x Reaction buffer (ph 9.0): Tris-HCl, KCl, etc. 1 x Dilution buffer (ph 8.0): Tris-HCl, EDTA, DTT, stabilizers, etc. dntps mixture: 10 mm, each dntp 2.5 mm Description Top DNA Polymerase 는 Thermophile polymerase gene 을 DNA recombination 기술을이용하여 polymerase activity 를향상시킨후 cloning 하여, E. coli 에발현및고순도로정제한 94 kda 의 thermostable DNA polymerase 입니다. Top DNA polymerase 의 quality 는 activity test, SDS-PAGE / purity test, endonuclease / nickase test 를수행하여확인하였고, PCR 방법을사용한다양한 DNA 증폭실험에사용할수있습니다. Features and Benefits High yields & High sensitivity: Taq DNA polymerase에비해향상된증폭효율과민감도를갖습니다. Stable reaction: 최적화된 buffer를제공하여 Top DNA polymerase의안정적반응을보증합니다. Reproducibility: 재현성있는결과를위해바이오니아의전제품은엄격한 ISO 품질시스템하에서생산됩니다. Concentration 500 U (5 U/μl) Storage Conditions ph 8.0, Tris-HCl, EDTA, DTT, stabilizers, etc. Storage Temperature - 20 Unit Definition One unit is defined at the amount of enzyme that will incorporate 10 nmole of dntp into acid-insoluble material in 30 min at 72. Specifications 5 to 3 exonuclease activity: No 3 to 5 exonuclease activity: No 3 -A overhang: Yes Fragment Size: ~10 kb www.bioneer.co.kr 31

Top DNA Polymerase Experimental Data Figure 1. Enzyme activity test of Top DNA Polymerase and Taq DNA polymerase. Top DNA polymerase/taq DNA Polymerase was serially diluted and used to amplify 20 ng of each lambda and human genomic DNA. Lane 1: 1 U of Top DNA Polymerase used Lane 2: 0.5 U of Top DNA Polymerase used Lane 3: 0.33 U of Top DNA Polymerase used Lane 4: 0.25 U of Top DNA Polymerase used Lane 5: 1 U of Taq DNA Polymerase used Lane 6: 0.5 U of Taq DNA Polymerase used Lane 7: 0.33 U of Taq DNA Polymerase used Lane 8: 0.25 U of Taq DNA Polymerase used Lane M:100 bp plus DNA Ladder (Bioneer, Cat. no. D-1035) Figure 2. Long PCR amplification test of Top DNA Polymerase and Taq DNA Polymerase using Lambda DNA. 10 ng of Lambda DNA and 1 U of each DNA Polymerase used for amplification. Lane 1: 2 kb PCR product Lane 2: 3 kb PCR product Lane 3: 4 kb PCR product Lane 4: 5 kb PCR product Lane 5: 6 kb PCR product Lane 6: 7 kb PCR product Lane 7: 8 kb PCR product Lane 8: 9 kb PCR product Lane 9: 10 kb PCR product M1: 1 kb DNA Ladder (Bioneer, Cat. no. D-1040) M2: Lambda DNA/Hind lll Marker (Bioneer, Cat. no. D-1050) Note: 본제품에포함된 Top DNA Polymerase 는 base incorporation rate 를증가시키기위하여 point mutation 을통해 5 ->3 exonuclease 활성을제거한효소입니다. 그러므로 SYBR green 을이용한 Real-Time qpcr 에는사용하실수있지만, TaqMan probe 등의 probe 를이용하는 Real- Time PCR 에는사용하실수없습니다. TaqMan probe 를이용한 qpcr 실험을위하여 AccuPower Dualstar TM qpcr PreMix 제품을권장합니다. Ordering Information Cat. no. Product Description D-3001 10 mm dntps Mix (each 2.5 mm), 1 ml E-3100 Top DNA Polymerase, 500 U, 10 mm dntps, 10 x reaction buffer with MgCl 2 E-3100-1 Top DNA Polymerase, 500 U, 10 mm dntps, 10 x reaction buffer, 20 mm MgCl 2 E-3100-2 Top DNA Polymerase, 500 U, 10 x reaction buffer with MgCl 2 E-3100-3 Top DNA Polymerase, 500 U, 10 x reaction buffer, 20 mm MgCl 2 E-3101 Top DNA Polymerase, 2,000 U, 10 mm dntps, 10 x reaction buffer with MgCl 2 E-3101-1 Top DNA Polymerase, 2,000 U, 10 mm dntps, 10 x reaction buffer, 20 mm MgCl 2 E-3101-2 Top DNA Polymerase, 2,000 U, 10 x reaction buffer with MgCl 2 E-3101-3 Top DNA Polymerase, 2,000 U, 10 x reaction buffer, 20 mm MgCl 2 32

Taq DNA Polymerase Versatile DNA Polymerase for Everyday Routine PCR Application Real-Time quantification of DNA and cdna targets using Dual probe, SYBR Green dye Gene expression profiling Microbial & viral pathogen detection Reagents Supplied 10 x Reaction buffer (ph 9.0): Tris-HCl, KCl, etc. Dilution buffer (ph 8.0): Tris-HCl, KCl, EDTA, DTT, stabilizers, etc. dntps mixture: 10 mm, each dntp 2.5 mm Description Taq DNA Polymerase 는 Thermus aquaticus YT1 유래 DNA polymerase gene 을재조합기술을이용하여 Escherichia coli 로부터발현시킨후순수분리정제된 94 kda 의내열성 Thermus aquaticus DNA polymerase 입니다. Features and Benefits High yields & High sensitivity: 높은증폭효율과 template 에대한높은민감도를갖습니다. Equipment Compatibility: 여러종류의실시간 PCR 장비에최적화된결과를얻을수있습니다. Concentration 500 U (5 U/μl) Storage Conditions ph 8.0, Tris-HCl, KCl, EDTA, DTT, stabilizers, etc. Storage Temperature -20 Unit Definition One unit is defined at the amount of enzyme that will incorporate 10 nmole of dntp into acid-insoluble material in 30 min at 72. Stable Reaction: 최적화된버퍼를제공하여 Taq DNA Polymerase 의안정적반응을보증합니다. Reproducibility: 재현성있는결과를위해바이오니아의전제품은엄격한 ISO 품질시스템하에서생산됩니다. Specifications 5 to 3 exonuclease activity: Yes 3 to 5 exonuclease activity: No 3 - A overhang: Yes Fragment size: ~10 kb www.bioneer.co.kr 33

Taq DNA Polymerase Experimental Data Figure 1. Using 1 unit of Taq DNA Polymerase, the activity of the polymerase was tested on human genomic DNA (A), lambda genomic DNA (B) as template. Each template DNA was serially diluted by tenfolds, with different ranges. Lane 1: 100 ng Template DNA Lane 2: 10 ng Template DNA Lane 3: 1 ng Template DNA Lane 4: 100 pg Template DNA Lane 5: 10 pg Template DNA Lane 6: 1 pg Template DNA Lane 7: 100 fg Template DNA M: 1 kb DNA Ladder (Bioneer, Cat. no. D-1040) Figure 2. Amplification of fragments ranging from 5 kb to 8 kb from template Lambda DNA 20 pg with 1 units of Taq DNA Polymerase. Lane 1: 5 kb PCR product Lane 2: 6 kb PCR product Lane 3: 7 kb PCR product Lane 4: 8 kb PCR product M: 1 kb DNA Ladder (Bioneer, Cat. no. D-1040) Ordering Information Cat. no. Product Description D-3001 10 mm dntps Mix (each 2.5 mm), 1 ml E-2011 Taq DNA Polymerase, 500 U, 10 mm dntps, 10 x reaction buffer with MgCl 2 E-2011-1 Taq DNA Polymerase, 500 U, 10 mm dntps, 10 x reaction buffer, 20 mm MgCl 2 E-2011-2 Taq DNA Polymerase, 500 U, 10 x reaction buffer with MgCl 2 E-2011-3 Taq DNA Polymerase, 500 U, 10 x reaction buffer, 20 mm MgCl 2 E-2013 Taq DNA Polymerase, 2,000 U, 10 mm dntps, 10 x reaction buffer with MgCl 2 E-2013-1 Taq DNA Polymerase, 2,000 U, 10 mm dntps, 10 x reaction buffer, 20 mm MgCl 2 E-2013-2 Taq DNA Polymerase, 2,000 U, 10 x reaction buffer with MgCl 2 E-2013-3 Taq DNA Polymerase, 2,000 U, 10 x reaction buffer, 20 mm MgCl 2 34

Pfu DNA Polymerase Novel Enzyme for High Fidelity PCR with DNA Proofreading Specifications 5 to 3 exonuclease activity: No 3 to 5 exonuclease activity: Yes 3 -A overhang: No Fragment size: ~10 kb Application Gene synthesis PCR or Primer extension requested high fidelity Blunt-end PCR Cloning or mutagenesis requested high fidelity Description Pfu DNA Polymerase 는 Pyrococus furiosus 라는 bacteria 에서유래되었으며, 3 5 exonuclease (proofreading) activity 를지니고있어기존의 Taq DNA Polymerase 보다 heat stability 와 fidelity 가뛰어납니다. 또한뛰어난 specificity 를가지고있어 nonspecific product 의발생이적으며, proofreading 기능이있어 DNA 증폭시발생하는 error rate 을감소시킵니다. Pfu DNA Polymerase 는 gene cloning, PCR, primer extension 등다양한실험에사용하실수있습니다. Features and Benefits High Fidelity PCR: 3 5 proofreading activity 가뛰어납니다. Thermostability: 열안정성이뛰어나 95 에서 1 시간반응후에도 94~99% 의기능을유지합니다. Terminal Transferase Activity: Terminal transferase activity 가없어 blunt-ended PCR product 를얻을수있습니다. Reagents Supplied 10 x Reaction Buffer (ph 9.0): Tris-HCl, KCl, etc. Dilution buffer (ph 8.0): Tris-HCl, EDTA, DTT, Stablizers, etc. dntps mixture: 10 mm, each dntp 2.5 mm (optional) Concentration 250 U (2.5 U/μl) Storage Conditions ph 8.0, Tris-HCl, EDTA, DTT, stabilizers, etc. Storage Temperature -20 Unit Definition One unit is defined at the amount of enzyme that will incorporate 10 nmol of dntp into acid-insoluble material in 30 min at 72. Reproducibility: 재현성있는결과를위해바이오니아의전제품은엄격한 ISO 품질시스템하에서생산됩니다. www.bioneer.co.kr 35

Pfu DNA Polymerase Experimental Data Figure 1. Human DNA was amplified using 2.5 units of enzyme in 50 μl reaction volume. Lane 1: 20 ng Lane 2: 2 ng Lane 3: 200 pg Lane 4: 20 pg M: 100 bp DNA ladder (Bioneer, Cat. no. D-1030) Figure 2. Lambda DNA 를이용한 Pfu DNA Polymerase 의 Long kb PCR test. Lane 1: Lambda DNA 5 kb Lane 2: Lambda DNA 6 kb Lane 3: Lambda DNA 7 kb Lane 4: Lambda DNA 8 kb M: 1 kb DNA ladder (Bioneer, Cat. no. D-1040) Ordering Information Cat. no. Product Description E-2015 Pfu DNA Polymerase, 250 U, 10 x reaction buffer E-2015-1 Pfu DNA Polymerase, 250 U, 10 mm dntps, 10 x reaction buffer E-2016 Pfu DNA Polymerase, 1,000 U, 10 x reaction buffer 36

ProFi Taq DNA Polymerase For High Efficiency and Amplification of Long Range PCR. Specifications 5 to 3 exonuclease activity: Yes 3 to 5 exonuclease activity: Yes 3 -A overhang: Yes Fragment size: ~30 kb Application Primer extension long-range amplification from genomic DNA High amplification efficiency Excellent performance on difficult templates Amplification of low-copy targets High yield and high sensitivity PCR Description ProFi Taq DNA Polymerase 는합성생물학기술을이용하여개발한 recombinant Taq DNA polymerase 로써뛰어난증폭효율과정확성을동시에충족시키는 DNA polymerase 입니다. ProFi Taq DNA Polymerase 는 human genomic DNA 의경우 21 kb 까지 DNA fragment 증폭이가능하며 lambda DNA 의경우 30 kb 까지 DNA fragment 증폭이가능합니다. ProFi Taq DNA Polymerase 는 high efficiency 및 long range PCR 에매우적합한제품이며그외에도 complex genomic DNA 또는 cdna templates, low-copy targets 등다양한 PCR 증폭반응에사용하실수있습니다. Features and Benefits High Sensitivity: 뛰어난민감도와증폭효율로미량의 human gdna target을검출합니다. 타사제품과의비교실험을통해우수한민감도및증폭효율을가집니다. Long Range PCR: Lambda DNA의경우 30 kb, human genomic DNA의경우 21 kb의 PCR 산물을효과적으로증폭할수있습니다. Reproducibility: 재현성있는결과를위해바이오니아의전제품은엄격한 ISO 품질시스템하에서생산됩니다. Reagents Supplied 10 x Reaction Buffer (ph 9.0): Tris-HCl, KCl, etc. Dilution buffer (ph 8.0): Tris-HCl, EDTA, DTT, KCl, Stablizers, etc. dntps mixture: 10 mm, each dntp 2.5 mm Concentration 250 U (5 U/μl) Storage Condition ph 8.0, Tris-HCl, KCl, EDTA, DTT, stabilizers, etc. Storage Temperature -20 www.bioneer.co.kr 37

ProFi Taq DNA Polymerase Experimental data Figure 1. Comparison of PCR amplification efficiency between ProFi Taq DNA Polymerase from Bioneer and other suppliers DNA polymerase. The cycling conditions for ProFi Taq DNA Polymerase were 95 for 5 min, 30 cycles of 95 for 20 sec, 55 for 20 sec and 72 for 30 sec. PCR reaction using other suppliers DNA polymerase were performed according to each supplier s protocol. Target: human Insulin receptor gene Lane 1: 10 ng of human genomic DNA Lane 2: 1 ng of human genomic DNA Lane 3: 100 pg of human genomic DNA Lane 4: 10 pg of human genomic DNA Lane M: 100 bp DNA Ladder(Bioneer, Cat. no. D-1030) Figure 2. Comparison of PCR amplification of long targets between ProFi Taq DNA Polymerase from Bioneer and other suppliers DNA polymerase The cycling conditions for ProFi Taq DNA Polymerase were 95 for 5 min, 32 cycles of 95 for 20 sec and 68 for 15 min. PCR reactions using other suppliers DNA polymerase were performed according to each supplier s protocol. Human genomic DNA was used as a template for PCR amplification. Lane 1: 11 kb fragment Lane 2: 13.5 kb fragment Lane 3: 17.6 kb fragment Lane 4: 21.4 kb fragment Lane M1: Lambda/Hind III marker (Bioneer, Cat. no. D-1050) Lane M2: 1 kb DNA Ladder (Bioneer, Cat. no. D-1040) Ordering Information Cat. no. Product Description E-2201 ProFi Taq DNA Polymerase 250 U, 10 mm dntps, 10 X reaction buffer with MgCl2 E-2202 ProFi Taq DNA Polymerase 250 U, 10 mm dntps, 10 X reaction buffer without MgCl2, 20 mm MgCl2 E-2203 ProFi Taq DNA Polymerase 250 U, 10 X reaction buffer with MgCl2 E-2204 ProFi Taq DNA Polymerase 250 U, 10 X reaction buffer without MgCl2, 20 mm MgCl2 E-2205 ProFi Taq DNA Polymerase 1000 U, 10 mm dntps, 10 X reaction buffer with MgCl2 E-2206 ProFi Taq DNA Polymerase 1000 U, 10 mm dntps, 10 X reaction buffer without MgCl2, 20 mm MgCl2 E-2207 ProFi Taq DNA Polymerase 1000 U, 10 X reaction buffer with MgCl2 E-2208 ProFi Taq DNA Polymerase 1000 U, 10 X reaction buffer without MgCl2, 20 mm MgCl2 38

HotStart DNA Polymerase Unique Enzyme-Mediated HotStart DNA Polymerase Features and Benefits High Sensitivity & High Specificity: 상온에서의 DNA polymerase 활성을완벽하게억제하였기때문에 PCR 반응물을혼합하는동안형성될수있는 nonspecific band나 primer-dimer의생성을억제합니다. Improve product yields: 높은 sensitivity 와 specificity 로인해원하는증폭산물의 yield 를향상시킬수있습니다. Reproducibility: 재현성있는결과를위해바이오니아의전제품은엄격한 ISO 품질시스템하에서생산됩니다. Description 항체를사용하여 polymerase 를불활성화시켰던기존의 Hot- Start 방법과는달리, PCR 반응에필수적인 2 가양이온 (Mg 2+ ) 을불활성화시키는 pyrophosphate (PPi) 와 70 에서활성을갖는내열성 pyrophosphatase (PPase) 를사용한신개념의 Hot- Start DNA polymerase 입니다. PCR 반응의 General Equation Template + Primer + Mg 2+ +dntp Elongation + 2Pi 일반적인 PCR 반응성분중에서 Mg 2+ 는 DNA polymerase의 activity에중요한역할을합니다. Mg 2+ 와결합력이강한 PPi를이용하여 PCR 초기단계에 Mg 2+ 의기능을억제하면비특이적증폭을최소화할수있습니다. PCR 반응액에 PPase와 PPi를넣어주면 PPi는 Mg 2+ 와결합하여 Mg 2+ -PPi complex를형성하고 PCR 반응을억제함으로써 PCR 초기단계에서발생하는비특이반응을방지합니다. 그후PCR 반응액이 70 이상이되면 PPase가활성화되어 PPi가 2Pi로분해되고 Mg 2+ 와해리됨으로써정상적인 PCR 반응이진행됩니다. 또한반응부산물이면서동시에 PCR 저해제인 PPi를 PPase가 2Pi 로분해함으로써 PCR 반응성을증가시킵니다. 무엇보다항체를사용하지않기때문에항체제거 step이필요치않아 PCR 반응시간을단축시키는장점을가지고있습니다. Specifications 5 to 3 exonuclease activity: No 3 to 5 exonuclease activity: No 3 -A overhang: Yes Fragment size: ~12 kb Application HotStart PCR, PCR with complex genomic templates/low copy templates/cdna Multiplex PCR Primer extension SNP typing Real-Time PCR using SYBR Green dye Multiple primer pairs and amplification of low copy template DNA Reagents Supplied 10 x Reaction buffer (ph 9.0): Tris-HCl, KCl, and Pyrophosphate 1 x Dilution buffer (ph 8.2): Tris-HCl, EDTA, DTT, stabilizers, etc. dntps mixture: 10 mm, each dntp 2.5 mm 20 mm MgCl 2 Concentration 250 U (5 U/μl) www.bioneer.co.kr 39

HotStart DNA Polymerase Storage Conditions ph 8.2, Tris-HCl, KCl, EDTA, DTT, stabilizers, etc. Storage Temperature -20 Unit Definition One unit is defined at the amount of enzyme that will incorporate 10 nmole of dntp into acid-insoluble material in 30 min at 72. Experimental Data Figure 1. Multiplex PCR comparison of genomic DNA using 6 sets of primers and 2 different DNA polymerases. Lane 1: 750 bp fragment Lane 2: 590 bp fragment Lane 3: 450 bp fragment Lane 4: 360 bp fragment Lane 5: 260 bp fragment Lane 6: 150 bp fragment Lane 7: Multiplex PCR with primers used for Lane 1~6 Lane M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030) Ordering Information Cat. no. Product Description E-3150 HotStart DNA Polymerase, 250 U, 10 mm dntps, 10 X reaction buffer 20 mm MgCl2 E-3150-1 HotStart DNA Polymerase, 250 U, 10 X reaction buffer 20 mm MgCl2 E-3151 HotStart DNA Polymerase, 1,000 U, 10 mm dntps, 10 X reaction buffer 20 mm MgCl2 40

HotStart Taq DNA Polymerase For Increased Specificity and Robust Sensitivity. Application Real-Time quantification of DNA and cdna targets using SYBR Green dye. HotStart PCR Multiplex PCR Automated PCR Reagents Supplied 10X Reaction Buffer (ph 9,0): Tris-HCl, KCl, etc. Dilution Buffer (ph 8.0): Tris-HCl, KCl, EDTA, DTT, Stabilizers, etc. dntps mixture: 10 mm, each dntp 2.5 mm Description HotStart Taq DNA Polymerase 는상온에서는활성이없다가 95 에서활성을가지게함으로써특이성이높은 PCR 산물을만들어낼수있도록개발되었습니다. 일반적인 PCR 반응시상온에서 PCR 반응물들을섞을때부터발생하는 mispriming 이나 primer 의 dimer 형성에의한비특이적인증폭을방지할수있습니다. Features and Benefits High Specificity & Fidelity: 일반적인 Taq DNA polymerase나 PCR premix보다우수한결과를얻을수있습니다. High Sensitivity: 불필요한반응이감소됨으로써소량의 template 으로도 PCR 수행이가능합니다. Concentration 250 U (5 U/μl) Storage Conditions ph 8.0, Tris-HCl, KCl, EDTA, DTT, Stabilizers, etc. Storage Temperature -20 Unit Definition One unit is defined at the amount of enzyme that will incorporate 10 nmole of dntp into acid-insoluble material in 30 min at 72. Experimental Data Easy Handling: 상온에서 polymerase 가활성을띄지않으므로 PCR 반응물을섞을때비특이적 priming 을방지할수있습니다. Simplicity: PCR 산물은 3 말단에 A tail 을가지고있어 T vector 에바로 cloning 할수있습니다. Reproducibility: 재현성있는결과를위해바이오니아의전제품은엄격한 ISO 품질시스템하에서생산됩니다. Specifications 5 to 3 exonuclease activity: Yes 3 to 5 exonuclease activity: No 3 -A overhang: Yes Fragment size: ~10 kb Figure 1. Specificity comparison between standard Taq and HotStart Taq DNA Polymerase. Single and multiples PCR results in human genomic DNA p53 Gene amplification Lane 1: 139 bp Lane 2: 211 bp Lane 3: 447 bp Lane 4: 618 bp Lane 5: 1,082 bp Lane 6: 1,296 bp Lane 7: 1,561 bp Lane 8: Multiplex PCR (139 bp, 447 bp, 618 bp) M: 100 bp DNA Ladder (Bioneer, Cat. no D-1030) www.bioneer.co.kr 41

HotStart Taq DNA Polymerase Figure 2. Performance comparison between HotStart Taq Polymerase and other supplier. M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1040) A: competitor A B: competitor B Ordering Information Cat. no. Product Description E-2017 HotStart Taq DNA Polymerase, 250 U, 10 mm dntps, 10 x reaction buffer with MgCl 2 E-2017-1 HotStart Taq DNA Polymerase, 1,000 U, 10 mm dntps, 10 x reaction buffer with MgCl 2 E-2017-2 HotStart Taq DNA Polymerase, 500 U, 10 mm dntps, 10 x reaction buffer with MgCl 2 E-2017-3 HotStart Taq DNA Polymerase, 250 U, 10 x reaction buffer with MgCl 2 E-2017-4 HotStart Taq DNA Polymerase, 1,000 U, 10 x reaction buffer with MgCl 2 42

02 RNA Amplification Standard RT & RT-PCR Kit AccuPower RT PreMix 44 AccuPower RT-PCR PreMix 46 High Efficiency RT Kit AccuPower CycleScript RT PreMix 48 High Sensitivity RT & RT-PCR Kit AccuPower RocketScript RT PreMix 51 AccuPower RocketScript Cycle RT PreMix 53 AccuPower RocketScript RT-PCR PreMix 56 Long & High Efficiency RT Kit AccuPower RocketScript RT PreMix, RNase H Minus New 58 AccuPower RocketScript RT-PCR PreMix, RNase H Minus New 60 Multiplex RT-PCR Kit AccuPower RocketPlex RT-PCR PreMix 61 HotStart RT & RT-PCR Kit AccuPower Dual-HotStart RT-PCR PreMix 64 AccuPower Dual-HotStart RT-PCR PreMix (with UDG) 66 Reverse Transcriptase M-MLV Reverse Transcriptase 68 CycleScript Reverse Transcriptase 69 RocketScript TM Reverse Transcriptase 72 RocketScript TM Reverse Transcriptase, RNase H Minus 74 Conventional PCR Instrument AllInOneCycler Go to M. Instruments & Devices

AccuPower RT PreMix M-MLV RTase 기반의표준 cdna 합성 Application First-strand synthesis of cdna from RNA molecules (RT) RT-PCR Random priming reaction cdna library construction Probe labeling mrna 5 end mapping by primer extension analysis Real-Time PCR Storage Temperature -20 Description AccuPower RT PreMix 는 RNA 를 template 로하여 cdna 를합성할때, reverse transcriptase, RNase inhibitor 등 cdna 합성에필요한구성물질들을 PreMix type 으로건조시킨제품입니다. 따라서사용자는 RNA, primer 및 D.W. 만을첨가하여 RT 반응을수행할수있습니다. 또한, 각 tube 에 1 회사용분량이분주된 single use 제품으로, 반복사용을하는 master mix 제품에서발생하는 cross contamination 문제를방지합니다. Experimental Data Sensitivity & Reproducibility Features and Benefits Ease-of-use: 각각의 PCR tube 에 M-MLV RTase 와 cdna 합성에필요한모든구성성분이포함되어있어 template RNA, primer, D.W. 만넣어바로 RT 반응을수행할수있습니다. cdna 합성이끝난반응물은 AccuPower PCR PreMix 을이용하여곧바로 PCR 반응을수행할수있습니다. Stability: RT 반응혼합물에안정화제를첨가하여건조시켜, 장기간보관하더라도활성을안정적으로유지합니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. Reliability Test: 모든 template RNA 로부터정확하게 cdna 합성이가능하며재현성있는결과를확인할수있습니다. Figure 1. Amplification of GAPDH target gene was detected using human total RNA (from 10 ng to 10 pg) with AccuPower RT PreMix. Template RNA: Total RNA from HeLa cells Lane 1: 10 ng Lane 2: 1 ng Lane 3: 100 pg Lane 4: 10 pg Con. Batch 1 Batch 2 Batch 3 Batch 4 Batch 5 Batch 6 10 pg 32.27 32.43 32.47 32.30 31.96 32.20 100 pg 28.78 28.39 28.73 29.12 29.01 29.04 1 ng 25.32 24.95 25.36 25.62 25.52 25.68 10 ng 22.04 21.68 22.11 22.51 22.09 22.37 Specifications Enzyme: M-MLV RTase DNase activity: No RNase activity: No RNase H activity: Yes Fragment size: ~ 9 kb 44

AccuPower RT PreMix Specific Amplification 1 2 3 4 5 Reliability and Reproducibility Test M 1 2 3 4 M 1 2 3 4 M 1 2 3 4 Figure 2. Specific amplification of 5 - UTR region of HCV with AccuPower RT PreMix. Lane 1: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030) Lane 2: negative control Lane 3: HCV positive serum Lane 4: HCV positive serum Lane 5: HCV negative serum Figure 3. AccuPower RT PreMix from each lot was tested to confirm reliability and reproducibility. Human total RNA (panel A: actin, panel B: globin) and hog cholera virus RNA (panel C) were used as template. Following cdna synthesis, AccuPower PCR PreMix was used to amplify target genes. Lane 1-4: Reliability test of each lot with AccuPower RT PreMix Lane M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030) Ordering Information Cat. no. Product Description K-2040 AccuPower RT PreMix, 0.5 ml thin-wall tubes with attached cap / 100 tubes, 20 μl rxn K-2041 AccuPower RT PreMix (-dye), 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 20 µl rxn K-2041-B AccuPower RT PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 20 μl rxn K-2042 AccuPower RT PreMix, 0.5 ml thin-wall tubes with attached cap / 100 tubes, 50 μl rxn K-2043 AccuPower RT PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 50 μl rxn K-2043-B AccuPower RT PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 50 μl rxn K-2082-1 AccuPower RT PreMix, thin-wall 384-well full-skirted plate, 5 μl rxn K-2082-2 AccuPower RT PreMix, thin-wall 384-well full-skirted plate, 10 μl rxn K-2082-3 AccuPower RT PreMix, thin-wall 384-well full-skirted plate, 20 μl rxn K-2261-1 AccuPower RT PreMix, thin-wall 96-well flat plate, 10 μl rxn K-2261-2 AccuPower RT PreMix, thin-wall 96-well full-skirted plate, 10 μl rxn K-2261-3 AccuPower RT PreMix, thin-wall 96-well semi-skirted plate, 10 μl rxn K-2261-4 AccuPower RT PreMix, thin-wall 96-well flat plate, 20 μl rxn K-2261-5 AccuPower RT PreMix, thin-wall 96-well full-skirted plate, 20 μl rxn K-2261-6 AccuPower RT PreMix, thin-wall 96-well semi-skirted plate, 20 μl rxn www.bioneer.co.kr 45

AccuPower RT-PCR PreMix M-MLV RTase 와 Top DNA Polymerase 기반의표준 One-Step RT-PCR Specifications Enzyme: M-MLV RTase, Top DNA polymerase 5 to 3 exonuclease: No 3 to 5 exonuclease: No 3 - A overhang: Yes RNase H activity: Yes Fragment size: ~ 5 kb Application First-strand synthesis of cdna from RNA molecules (RT) RT-PCR cdna library construction Gene expression analysis Description AccuPower RT-PCR PreMix 는 low-copy RNA (viral RNA) 나 mrna 를주형으로하는 cdna 합성과 PCR 의 2 단계반응을한 tube 내에서연속적으로수행함으로써간편하고경제적으로실험할수있는 one-step RT-PCR 제품입니다. RT-PCR 에사용되는모든시약을동결건조시키는바이오니아의특허기술을적용하여안정성이뛰어나며, 재현성이높은실험결과를얻을수있습니다. 또한, 각 tube 에 1 회사용분량이분주된 single use 제품으로, 반복사용을하는 master mix 제품에서발생하는 cross contamination 문제를방지합니다. Storage Temperature -20 Features and Benefits Ease-of-use: 각각의 PCR tube 에 M-MLV RTase,Top DNA polymerase 및 cdna 합성과 PCR 에필요한모든물질을포함하고있어 template RNA, primer set 와 D.W. 만넣어바로 one-step RT-PCR 반응을수행할수있습니다. 또한전기영동시에필요한 tracking dye 와침강제가포함되어있어 sample loading buffer 를첨가할필요가없으므로간편하게사용할수있습니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. Stability: AccuPower RT-PCR PreMix 는 PCR 반응혼합액에특수안정화물질을첨가하여건조함으로써장기보관하더라도활성을안정적으로유지합니다. Reliability Test: 모든 template RNA 로부터정확하게 RT-PCR 이가능하며재현성있는결과를확인할수있습니다. 46

AccuPower RT-PCR PreMix Experimental Data Sensitivity Reproducibility Batch1 Batch2 Batch3 Batch4 M 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 Figure 1. Sensitivity comparison between AccuPower RT-PCR PreMix and Supplier I RT-PCR kit. Each 10 9 copies ~10 3 copies of PSTVd used for RT/PCR and the same amount of RT/PCR products used for electrophoresis. Lane 1: 10 9 copy Lane 2: 10 8 copy Lane 3: 10 7 copy Lane 4: 10 6 copy Lane 5: 10 5 copy Lane 6: 10 4 copy Lane 7: 10 3 copy Lane 8: NTC Lane M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030) Figure 2. Comparison of reproducibility test for AccuPower RT- PCR PreMix batch 1, 2, 3 and batch 4 products using serial diluted human total RNA. Lane 1: 10 ng human total RNA from HeLa cell Lane 2: 1 ng human total RNA from HeLa cell Lane 3: 100 pg human total RNA from HeLa cell Lane 4: 10 pg human total RNA from HeLa cell Lane M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030) Ordering Information Cat. no. Product Description K-2055 AccuPower RT-PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 20 μl rxn K-2055-B AccuPower RT-PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 20 μl rxn K-2056 AccuPower RT-PCR PreMix, 0.5 ml thin-wall individual tubes with attached cap / 100 tubes, 50 μl rxn K-2057 AccuPower RT-PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 50 μl rxn K-2057-B AccuPower RT-PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 50 μl rxn K-2084-1 AccuPower RT-PCR PreMix, thin-wall 384-well full-skirted plate, 5 μl rxn K-2084-2 AccuPower RT-PCR PreMix, thin-wall 384-well full-skirted plate, 10 μl rxn K-2084-3 AccuPower RT-PCR PreMix, thin-wall 384-well full-skirted plate, 20 μl rxn K-2262-1 AccuPower RT-PCR PreMix, thin-wall 96-well flat plate, 10 μl rxn K-2262-2 AccuPower RT-PCR PreMix, thin-wall 96-well full-skirted plate, 10 μl rxn K-2262-3 AccuPower RT-PCR PreMix, thin-wall 96-well semi-skirted plate, 10 μl rxn K-2262-4 AccuPower RT-PCR PreMix, thin-wall 96-well flat plate, 20 μl rxn K-2262-5 AccuPower RT-PCR PreMix, thin-wall 96-well full-skirted plate, 20 μl rxn K-2262-6 AccuPower RT-PCR PreMix, thin-wall 96-well semi-skirted plate, 20 μl rxn www.bioneer.co.kr 47

AccuPower CycleScript RT PreMix (dt20 & dn12 & dn6) 순환온도역전사반응을통한고효율의 cdna 합성 Description AccuPower CycleScript RT PreMix (dt 20 & dn 12 & dn 6) 는순환온도역전사반응 (CTRT, Cyclic Temperature Reverse Transcription) 이적용된제품입니다. 세계적으로인정받은특허기술인순환온도역전사반응은합성효율을증가시킬뿐아니라 full-length cdna 합성에효과적입니다. 순환온도역전사반응은저온반응 (15~40 ) 에서 primer annealing 이진행되고고온 (50~55 ) 에서 template RNA 의 2 차구조형성을풀어주는 2~3 개 steps 을반복적으로적용하여기존의 42 역전사반응보다높은효율로 cdna 를합성할수있습니다. 반응에적합한농도의 oligo dt20, dn6, dn12 primer 가포함된제품을선택할수있어서사용자는 template RNA 만넣어손쉽게 cdna 합성을할수있습니다. 또한, 각 tube 에 1 회사용분량이분주된 single use 제품으로, 반복사용을하는 master mix 제품에서발생하는 cross contamination 문제를방지합니다. 상기단일온도역전사반응 (FTRT, Fixed Temperature Reverse Transcription) 및순환적역전사반응 (CTRT) 과정은다음과같습니다. 단일온도역전사반응 (FTRT) - Step 1: RNA denaturation 단계 (65, 10 min) - Step 2: cdna 합성단계 (42, 15 min) Features and Benefits Flexible Reaction Conditions: 순환온도역전사반응 (CTRT) 과더불어 42~55 내의단일온도에서도역전사반응 (FTRT) 이가능합니다. Stability: RT 반응혼합물에안정화제를첨가하여건조시켜, 장기간보관하더라도활성을안정적으로유지합니다. Controllable Reaction Time: 반응시간은증폭하려는유전자의 copy 수와크기에따라달라질수있습니다. High copy gene 의경우, 10 분의역전사반응으로도충분한양의 cdna 를합성할수있습니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. 순환온도역전사반응 (CTRT) - Step 1: Primer annealing 단계 (25~40, 30 sec) - Step 2: cdna 합성단계 (42~48, 4 min) -Step 3 (optional): DNA template 2 차구조해소및 cdna 합성단계 (50~55, 30 sec) * 상기반응에서온도, 시간및 cycle 수는실험자의실험조건에따라선택적으로설정할수있습니다. Ease-of-use: 각각의 PCR tube 에 CycleScript RTase 와 cdna 합성에필요한모든구성성분및 primer 가포함되어있어 template RNA, D.W. 만넣어바로 RT 반응을수행할수있습니다. cdna 합성이끝난반응물은 AccuPower PCR PreMix 을이용하여곧바로 PCR 반응을수행할수있습니다. Specifications Enzyme: CycleScript RTase DNase activity: No RNase activity: No RNase H activity: Yes Fragment size: ~ 9 kb 48

AccuPower CycleScript RT PreMix (dt20 & dn12 & dn6) Application Sequencing single and double-strand DNA or RNA RT-PCR Random priming reaction cdna Library construction Probe labeling mrna 5 -end mapping by primer extension analysis Real-Time PCR Storage Temperature -20 Experimental Data 2.6 kb 2.0 kb 1.5 kb 800bp 42 C 30 min 55 C 30 min (37 C 2 min/50 C 3min) X6 dt 20 dn 12 dt 20 dn 12 dt 20 dn 12 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 M 17 18 19 2021 22 23 24 Figure 1. Reaction conditions of CTRT compared with that of conventional RT. The conventional fixed temperature RT reactions at 42 and 55 and cyclic temperature RT reactions at 37 and 50 using HeLa cell total RNA (800 ng each) were performed 800 bp, 1.5 kb, 2.0 kb, and 2.6 kb fragments of human transferrin receptor gene were amplified with each primers. AccuPower CycleScript RT PreMix series (dt20 & dn12) preferably shows good results at CTRT reaction condition. 3.5 kb 2.6 kb 2.0 kb 1.5 kb Bioneer-dT Bioneer-random Supplier C-dT Supplier C-dN Supplier I-dT Supplier I-dN 1 2 3 4 5 6 7 8 9 10 M 11 12 13 14 15 16 17 18 19 20 M 212223 24 25 26 2728 29 30 Figure 3. RT reaction at various temperature and short reaction time. Rxn. condition: conventional 42 1 hr, 55 1hr & cyclic reaction 1: (37 2 min/50 3 min)x 12, cyclic reaction 2: (37 1 min/ 47 3 min/ 55 1 min) X 12. This product shows thermal stability. Target: Human GAPDH, human β-actin primer set: human GAPDH primer set human β-actin primer set Lane 1, 5, 9, 13, 17, 21, 25 & 29: HeLa cell total RNA 100 ng Lane 2, 6, 10, 14, 18, 22, 26 & 30: HeLa cell total RNA 10 ng Lane 3, 7, 11, 15, 19, 23, 27 & 31: HeLa cell total RNA 1 ng Lane 4, 8, 12, 16, 20, 24, 28 & 32: HeLa cell total RNA 100 pg Lane M: 100 bp DNA Ladder (Bioneer, Cat. no. D-1030) Rxn. time : 60min 20min 10min 60min 20min 10min 1 2 3 4 M 5 6 7 8 9 10 1112 1314 15 16 M 17 1819 20 21 22 23 24 1 kb CAPDH 300 bp actin Figure 4. Amplification results at various reaction times. CycleScript RT PreMix (dt20) was performed cyclic temperature RT according to (37 2 min/50 3 min); X12, X4, X2 cycles. 10 min reaction is enough. HeLa cell total RNA templates were serially diluted such as 100 ng, 10 ng, 1 ng, 100 pg. 800 kb Figure 2. Gene amplification test of transferrin receptor compared with other companies. The reaction condition was performed according to each manufacturer s recommendation. All cdnas were reacted with AccuPower PCR PreMix (Bioneer, Cat. no. K-2012). Lane 1~5: AccuPower CycleScript RT PreMix (dt20) incubated at 55 for 1 hr Lane 6~10: AccuPower CycleScript RT PreMix (dn12) incubated at 55 for 1 hr Lane 11~15: C company s RT product including dt primer incubated at 42 for 1 hr Lane 16~20: C company s RT product including random primer incubated at 42 for 1 hr Lane 21~25: I company s RT product including dt primer incubated at 45 for 1 hr Lane 26~30: I company s RT product including random primer incubated at 45 for 1 hr Lane M: 1 kb DNA Ladder (Bioneer, Cat. no. D-1040) www.bioneer.co.kr 49

AccuPower CycleScript RT PreMix (dt20 & dn12 & dn6) Ordering Information Cat. no. Product Description K-2044 AccuPower CycleScript RT PreMix, dt 20 / 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 20 μl rxn K-2044-B AccuPower CycleScript RT PreMix, dt 20 / 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 20 μl rxn K-2045 AccuPower CycleScript RT PreMix dn 12/ 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 20 μl rxn K-2045-B AccuPower CycleScript RT PreMix, dn 12 / 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 20 μl rxn K-2046 AccuPower CycleScript RT PreMix, dn 6 / 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 20 μl rxn K-2046-B AccuPower CycleScript RT PreMix, dn 6 / 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 20 μl rxn K-2047 AccuPower CycleScript RT PreMix, dt 20 / 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 50 μl rxn K-2047-B AccuPower CycleScript RT PreMix, dt 20 / 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 50 μl rxn K-2048 AccuPower CycleScript RT PreMix, dn 12 / 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 50 μl rxn K-2048-B AccuPower CycleScript RT PreMix, dn 12 / 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 50 μl rxn K-2049 AccuPower CycleScript RT PreMix, dn 6 / 0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes, 50 μl rxn K-2049-B AccuPower CycleScript RT PreMix, dn 6 / 0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes, 50 μl rxn K-2050 AccuPower CycleScript RT PreMix, dt 20 / 0.5 ml thin-wall tubes with attached cap / 100 tubes, 50 μl K-2050-1 AccuPower CycleScript RT PreMix, dn 12 / 0.5 ml thin-wall individual tubes with attached cap / 100 tubes, 50 μl rxn K-2050-2 AccuPower CycleScript RT PreMix, dn 6 / 0.5 ml thin-wall individual tubes with attached cap / 100 tubes, 50 μl rxn 50

AccuPower RocketScript RT PreMix 고온안정성이뛰어난 RocketScript TM RTase 를적용하여 2 차구조 RNA 의 cdna 합성 Description AccuPower RocketScript TM RT PreMix 는바이오니아가독자적인기술로개발한 Thermostable RTase (RocketScript TM Reverse Transcriptase) 제품으로, 기존에어려웠던복잡한 2 차구조 RNA 의 cdna 합성을효과적으로수행할수있습니다. 일반적인 M-MLV RTase 는고온에서활성을잃어저온 (42 ) 에서 RT 를수행하므로 2 차구조 RNA 의 cdna 합성이어렵습니다. 열안정성을가진 RocketScript TM RTase 는고온 (70 ) 에서도활성을유지하여 2 차구조 RNA 를풀어내기때문에 full-length cdna 합성에용이합니다. 또한반응효율이높아 10 fg 의 human total RNA 도성공적으로역전사할수있습니다. 또한, 각 tube 에 1 회사용분량이분주된 single use 제품으로, 반복사용을하는 master mix 제품에서발생하는 cross contamination 문제를방지합니다. Features and Benefits Thermostable Activity: 70 에서도활성을갖는뛰어난열안정성의 RocketScript TM RTase 적용으로 2차구조 RNA의 cdna 합성에적합합니다. cdna 합성온도는 42~70 까지다양한온도로설정할수있어효과적으로 cdna를합성할수있습니다. Sensitivity: 민감도가뛰어나 100 fg의 human total RNA에서도효과적으로 cdna를합성할수있습니다. Ease-of-use: 각각의 PCR tube에 RocketScript TM RTase와 cdna 합성에필요한모든구성성분이포함되어있어 template RNA, primer, D.W. 만넣어바로 RT 반응을수행할수있습니다. cdna 합성이끝난반응물은 AccuPower PCR PreMix을이용하여곧바로 PCR 반응을수행할수있습니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system으로대량생산되어각 batch에대한철저한 QC를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. Specifications Enzyme: RocketScript TM RTase DNase activity: No RNase activity: No RNase H activity: Yes Fragment size: ~ 10 kb Note: Schematic representation of the 5 UTR of a gene, with complex secondary structure, at three different temperatures. Note that RocketScript TM RTase shows full activity at 70 allowing it to synthesize the complete gene sequence where M-MLV and other Reverse Transcriptase s fail. Application First-strand synthesis of cdna from RNA molecules (RT) RT-PCR Random priming reactions cdna library construction Probe labeling mrna 5 -end mapping by primer extension analysis Real-Time PCR Storage Temperature -20 www.bioneer.co.kr 51

AccuPower RocketScript RT PreMix Experimental Data Thermostable Activity A) Supplier I Reverse Transcriptase 42 1 2 3 4 5 6 7 8 50 1 2 3 4 5 6 7 8 M Enhanced Performance A) Supplier I Reverse Transcriptase (RT at 50 ) B) AccuPower RocketScript TM RT PreMix (RT at 50 ) B) AccuPower RocketScript TM RT PreMix 42 1 2 3 4 5 6 7 8 50 M 1 2 3 4 5 6 7 8 Figure 1. Sensitivity comparison between AccuPower RocketScript TM RT PreMix and M-MLV RTase. Sensitivity results of AccuPower RocketScript TM RT PreMix using GAPDH compared with conventional reverse transcriptases. Each 10 fg~100 ng of total RNA used for RT and the same amount of RT products used for electrophoresis. Lane 1: 10 fg human total RNA from HeLa cell Lane 2: 100 fg human total RNA from HeLa cell Lane 3: 1 pg human total RNA from HeLa cell Lane 4: 10 pg human total RNA from HeLa cell Lane 5: 100 pg human total RNA from HeLa cell Lane 6: 1 ng human total RNA from HeLa cell Lane 7: 10 ng human total RNA from HeLa cell Lane 8: 100 ng human total RNA from HeLa cell Figure 3. Comparison of amplification efficiency between Accu- Power RocketScript TM RT PreMix and competitors M-MLV RTase. RocketScript TM is able to handle a wide range of sample concentrations and transcript lengths so your downstream applications are minimally affected by the reverse transcription step. Lane 1: 1,000 ng of total RNA from HeLa cells Lane 2: 100 ng of total RNA from HeLa cells Lane 3: 10 ng of total RNA from HeLa cells Note: Competitor products show inhibition at high input concentrations of total RNA. Sensitivity Test Concentration RocktScript TM RT PreMix Supplier Q Supplier I 10,000 23.91 25.63 24.43 1,000 27.33 28.92 28.03 100 30.62 32.42 30.88 10 33.63 35.43 33.95 Efficiency 104% 103% 108% Linearity 0.9999 0.9996 0.9995 Figure 2. Sensitivity comparison between AccuPower RocketScript TM RT PreMix and Supplier RTases using Real-Time PCR. Rxn. conditions: conventional 1 hr incubation at 60, deactivation at 95 for 5 min All cdnas were amplified with AccuPower DualStar TM qpcr PreMix (Bioneer, Cat. no. K-6110). Ordering Information Cat. no. Product Description K-2101 AccuPower RocketScript TM RT PreMix, 20 μl, 0.2 ml thin-wall 8-tube strips with attached cap, 96 tubes K-2102 AccuPower RocketScript TM RT PreMix, 20 μl, 0.2 ml thin-wall 8-tube strips with attached cap, 480 tubes K-2103 AccuPower RocketScript TM RT PreMix, 50 μl, 0.2 ml thin-wall 8-tube strips with attached cap, 96 tubes K-2104 AccuPower RocketScript TM RT PreMix, 50 μl, 0.2 ml thin-wall 8-tube strips with attached cap, 480 tubes 52

AccuPower RocketScript Cycle RT PreMix 고온안정성이뛰어난 RocketScript TM RTase 와 CTRT 를적용하여고효율로 2 차구조 RNA 의 cdna 합성 Description AccuPower RocketScript TM Cycle RT PreMix 는바이오니아가독자적으로개발한 Thermostable RTase (RocketScript TM Reverse Transcriptase) 와순환온도역전사반응 (CTRT, Cyclic Temperature Reverse Transcription) 기술이융합된차세대 cdna synthesis 용제품입니다. 이제품은고온 (70 ) 에서도활성을유지할수있는 RocketScript TM RTase 를순환온도역전사반응 (CTRT) 에적합하도록최적화된제품으로, 안정한 2 차구조를가진 RNA 들도효과적으로 full-length cdna 로합성할수있습니다. 반응효율이높아 10 fg 의 human total RNA 에서도성공적으로 cdna 합성을할수있습니다. 반응에적합한농도의 Oligo dt20, dn6, dn12 primer 가포함된제품을선택할수있어서사용자는 template RNA 와 D.W. 만넣어손쉽게 cdna 합성을할수있습니다. 또한, 각 tube 에 1 회사용분량이분주된 single use 제품으로, 반복사용을하는 master mix 제품에서발생하는 cross contamination 문제를방지합니다. 효과적으로 cdna를합성할수있습니다. Flexible Reaction Conditions: 순환온도역전사반응 (CTRT) 과더불어 42~70 내의단일온도에서역전사반응 (FTRT, Fixed Temperature Reverse Transcription) 이가능합니다. Ease-of-use: 각각의 PCR tube에 RocketScript TM RTase와 cdna 합성에필요한모든구성성분이포함되어있어 template RNA 와 D.W. 만넣어바로 RT 반응을수행할수있습니다. cdna 합성이끝난반응물은 AccuPower PCR PreMix을이용하여곧바로 PCR 반응을수행할수있습니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system으로대량생산되어각 batch에대한철저한 QC를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. Specifications Enzyme: RocketScript TM RTase DNase activity: No RNase activity: No RNase H activity: Yes Fragment size: ~ 10 kb Application First-strand synthesis of cdna from RNA molecules (RT) RT-PCR Random priming reactions cdna library construction Probe labeling mrna 5 -end mapping by primer extension analysis Real-Time PCR Storage Temperature -20 Note: Schematic representation of the 5 UTR of a gene, with complex secondary structure, at three different temperatures. Note that RocketScript TM shows full activity at 70 allowing it to synthesize the complete gene sequence where M-MLV and other reverse transcriptase s fail. Features and Benefits Sensitivity & Efficiency: 순환온도역전사반응과 thermostable RTase (RocketScript TM RTase) 의융합기술은민감도와효율을증가시켜 cycle 수를조절하여낮은농도의 RNA로부터손쉽게 cdna를합성할수있습니다. Thermostable Activity: 70 에서도활성을갖는뛰어난열안정성의 RocketScript TM RTase 적용으로 2차구조 RNA의 cdna 합성에적합합니다. cdna 합성온도는 42~70 까지다양한온도로설정할수있어 www.bioneer.co.kr 53

AccuPower RocketScript Cycle RT PreMix Experimental Data Thermostable Activity Cyclic Temperature Reverse Transcription Figure 1. Complex RNA amplification results of AccuPower RocketScript TM Cycle RT PreMix. Each target gene (MYC, TFRC) was amplified after performing reverse transcription with AccuPower RocketScript TM Cycle RT PreMix. Rxn. conditions: Conventional 1 hr incubation at 42, 50, or 60, deactivation at 95 for 5 min A: M-MLV Reverse Transcriptase B: AccuPower RocketScript TM Cycle RT PreMix with Oligo (dt20) Lane 1: 100 ng Human total RNA from HeLa cell Lane 2: 10 ng Human total RNA from HeLa cell Sensitivity & Full-length cdna Synthesis Concentration (copies/rxn) FTRT (Ct) CTRT with 1 cycle (Ct) CTRT with 10 cycles (Ct) 10,000 19.46 18.77 18.51 1,000 24.11 24.04 22.93 100 29.78 28.35 28.19 10 32.87 33.00 31.05 Figure 3. Low copy species enrichment by cycle. Comparing fixed temperature RT to 1 and 10 cycle(s) of cyclic temperature RT reveal progressive improvement in detected cdna yield as input copies decrease FTRT: 60 min incubation at 50 followed by 5 min deactivation at 95 CTRT: Cycles of 37 annealing 10 sec, 50 cdna synthesis 4 min, 55 secondary structure melting and cdna synthesis 30 sec Target: Human GAPDH Human total RNA from HeLa cells qpcr with AccuPower GreenStar TM qpcr PreMix (Bioneer, Cat. no. K-6210) Figure 2. Comparison of amplification efficiency between AccuPower RocketScript TM Cycle RT PreMix and competitors RTases. (a) Sensitivity test Target: Human TFRC Lane 1: 100 ng human total RNA from HeLa cell Lane 2: 10 ng human total RNA from HeLa cell Lane 3: 1 ng human total RNA from HeLa cell Lane 4: 100 pg human total RNA from HeLa cell Lane M: 1 kb DNA Ladder (Bioneer, Cat. no. D-1040) RT reaction condition is performed according to each manufacturer s recommendations. (b) Full-Length cdna synthesis test RT reactions were performed according to each manufacturer s recommendation. All cdnas were amplified with AccuPower HotStart PCR PreMix (K-5050) from Bioneer Note: Competitor products show inhibition at high input concentrations of total RNA. Lane 1: 1 μg human total RNA from HeLa cell Lane 2: 100 ng human total RNA from HeLa cell Lane 3: 10 ng human total RNA from HeLa cell 54

AccuPower RocketScript Cycle RT PreMix Ordering Information Cat. no. Product Description K-2201 AccuPower RocketScript TM Cycle RT PreMix (dt 20), 20 μl, 0.2 ml thin-wall 8-tube strips with attached cap, 96 tubes K-2202 AccuPower RocketScript TM Cycle RT PreMix (dt 20), 20 μl, 0.2 ml thin-wall 8-tube strips with attached cap, 480 tubes K-2203 AccuPower RocketScript TM Cycle RT PreMix (dt 20), 50 μl, 0.2 ml thin-wall 8-tube strips with attached cap, 96 tubes K-2204 AccuPower RocketScript TM Cycle RT PreMix (dt 20), 50 μl, 0.2 ml thin-wall 8-tube strips with attached cap, 480 tubes K-2205 AccuPower RocketScript TM Cycle RT PreMix(dN 6), 20 μl, 0.2 ml thin-wall 8-tube strips with attached cap, 96 tubes K-2206 AccuPower RocketScript TM Cycle RT PreMix(dN 6), 20 μl, 0.2 ml thin-wall 8-tube strips with attached cap, 480 tubes K-2207 AccuPower RocketScript TM Cycle RT PreMix(dN 6), 50 μl, 0.2 ml thin-wall 8-tube strips with attached cap, 96 tubes K-2208 AccuPower RocketScript TM Cycle RT PreMix(dN 12), 20 μl, 0.2 ml thin-wall 8-tube strips with attached cap, 96 tubes K-2209 AccuPower RocketScript TM Cycle RT PreMix(dN 12), 20 μl, 0.2 ml thin-wall 8-tube strips with attached cap, 480 tubes www.bioneer.co.kr 55

AccuPower RocketScript RT-PCR PreMix 고온안정성이뛰어난 RocketScript TM RTase 를적용하여 2 차구조 RNA 의 One-step RT-PCR Features and Benefits Thermostable Activity: 70 에서도활성을갖는뛰어난열안정성의 RocketScript TM RTase 적용으로 2 차구조 RNA 의 cdna 합성에적합합니다. cdna 합성온도는 42~70 까지다양한온도로설정할수있어효과적으로 cdna 를합성할수있습니다. Sensitivity: AccuPower RocketScript TM RT-PCR PreMix 은 cell, tissue 등에서추출한 human total RNA 의 10 pg (target: ß-actin) 에서도 one-step RT-PCR 이가능한제품입니다. Description AccuPower RocketScript TM RT-PCR PreMix 는바이오니아가독자적으로개발한 thermostable RTase (RocketScript TM Reverse Transcriptase) 와 Top DNA polymerase, dntp, reaction buffer 등을동결건조시킨 one-step RT-PCR 제품입니다. cdna 합성과 PCR 의 2 단계반응을하나의 tube 에서 one-step 으로수행하는제품입니다. RocketScript TM Reverse Transcriptase 를이용하여고온에서 RT 반응을수행함으로써, RNA 2 차구조가강하게형성된 template RNA 에서도효과적으로 RT-PCR product 를얻을수있습니다. RocketScript TM RTase 는신장성이매우뛰어나 low copy RNA 로부터 cdna 를높은효율로합성할수있습니다. 또한, 각 tube 에 1 회사용분량이분주된 single use 제품으로, 반복사용을하는 master mix 제품에서발생하는 cross contamination 문제를방지합니다. Note: Schematic representation of the 5 UTR of a gene, with complex secondary structure, at three different temperatures. Note that RocketScript TM shows full activity at 70 allowing it to synthesize the complete gene sequence where M-MLV and other reverse transcriptase s fail. Long Range RT-PCR: Human total RNA 100 ng 으로 full-length size 가 5.2 kb (target: Human TFRC) target 도 one-step RT-PCR 이가능합니다. Ease-of-use: 각각의 PCR tube 에 RocketScript TM RTase, Top DNA polymerase 및 cdna 합성과 PCR 에필요한모든구성성분이포함되어있어 template RNA, primer set, D.W. 만을넣어바로 one-step RT-PCR 반응을수행할수있습니다. 또한전기영동시에필요한 tracking dye 와침강제가포함되어있어 sample loading buffer 를첨가할필요가없으므로간편하게사용할수있습니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. Specifications Enzyme: RocketScript TM RTase, Top DNA polymerase 5 to 3 exonuclease: No 3 to 5 exonuclease: No 3 - A overhang: Yes RNase H activity: Yes Fragment size: ~ 6 kb Application First-strand synthesis of cdna from RNA molecules (RT) RT-PCR cdna library construction Gene expression analysis Storage Temperature -20 56

AccuPower RocketScript RT-PCR PreMix Experimental Data Figure 1. Performance comparison between AccuPower RocketScript TM RT-PCR PreMix and competitor RT-PCR kits. Complex secondary structure RNA species amplification by RocketScript TM is outstanding compared to the leading competitor s reverse transcriptase. RT reactions were performed according to each manufacturer s recommendations. Lane 1: 10 ng of total RNA from HeLa cells Lane 2: 1 ng of total RNA from HeLa cells Lane 3: 100 pg of total RNA from HeLa cells Lane 4: 10 pg of total RNA from HeLa cells Figure 2. AccuPower RocketScript TM RT-PCR PreMix shows enhanced performance compared to competitors. One-step RT-PCR reactions were performed with 100 ng total RNA from HeLa cells using regents and conditions specified in each manufacturer s protocol. Lane 1: 3 kb Lane 2: 4.5 kb Lane 3: 5.2 kb RT reaction condition is performed according to each manufacturer s recommendations. Batch1 Batch2 Batch3 Batch4 M 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 Figure 3. Highly reproducible amplifications. Amplification of an 500 bp target gene was detected using human toal RNA (from 10 ng to 10 pg) with AccuPower RocketScript TM RT-PCR PreMix. As shown in figure 3, highly reproducible amplifications were achieved within each Lot. set of triplicates Lane 1: 10 ng of total RNA from HeLa cells Lane 2: 1 ng of total RNA from HeLa cells Lane 3: 100 pg of total RNA from HeLa cells Lane 4: 10 pg of total RNA from HeLa cells Ordering Information Cat. no. Product Description K-2501 AccuPower RocketScript TM RT-PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap/96 tubes 20 μl/rxn K-2503 AccuPower RocketScript TM RT-PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap/96 tubes 50 μl/rxn K-2502 AccuPower RocketScript TM RT-PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap/480 tubes 20 μl/rxn K-2504 AccuPower RocketScript TM RT-PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap/480 tubes, 50 μl/rxn www.bioneer.co.kr 57

AccuPower RocketScript RT PreMix, RNase H Minus NEW RNase H activity 를제거하여 Long RNA 의 Full-length cdna 합성 Ease-of-use: 각각의 PCR tube 에 RocketScript TM RTase 와 cdna 합성에필요한모든구성성분이포함되어있어 template RNA, primer, D.W. 만을넣어바로 RT 반응을수행할수있습니다. cdna 합성이끝난반응물은 AccuPower PCR PreMix 을이용하여곧바로 PCR 반응을수행할수있습니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. Description AccuPower RocketScript TM RT PreMix, RNase H Minus 는바이오니아에서독자적으로개발한 RocketScript TM RTase, RNase H Minus) 를사용한제품입니다. RNase H activity 는 cdna 합성시에 cdna 와 hybrid 된주형의 RNA 를제거하는역할을하는데너무강력한 RNase H 의활성을갖게되면 cdna 합성신장부위에도영향을주기때문에긴 size 의 cdna 를합성할수없게됩니다. AccuPower RocketScript TM RT PreMix, RNase H Minus 는독자적유전공학기술을이용하여 RNase H 활성을제거하여 long target RNA 의 cdna 합성에유용하며신장성및민감도가뛰어나 10 fg 의 human total RNA 에서도성공적으로 cdna 합성을할수있습니다. 또한 RocketScript TM Reverse Transcriptase (RNase H Minus), RNase inhibitor 등 cdna 합성에필요한물질들을반응 tube 에건조시켜 template RNA 및 primer, D.W. 의첨가만으로손쉽게 cdna 합성을할수있는편리한제품입니다. 또한, 각 tube 에 1 회사용분량이분주된 single use 제품으로, 반복사용을하는 master mix 제품에서발생하는 cross contamination 문제를방지합니다. Features and Benefits Thermostable Activity: 70 에서도활성을갖는뛰어난열안정성의 RocketScript TM RTase 적용으로 2 차구조 RNA 의 cdna 합성에적합합니다. cdna 합성온도는 42~70 까지다양한온도로설정할수있어효과적으로 cdna 를합성할수있습니다. Elimination RNase H activity: RocketScript TM RTase, RNase H Minus 는 cdna 합성과정에서생성되는 RNA/DNA hybrid 를분해하는 RNase H 활성이없어긴 target RNA 의 cdna 합성에적합하고높은민감도를가지므로낮은농도의 RNA 에서도향상된성능으로증폭할수있습니다. Sensitivity: 민감도가뛰어나 100 fg 의 human total RNA 에서도효과적으로 cdna 를합성할수있습니다. Specifications Enzyme: RocketScript TM RTase DNase activity: No RNase activity: No RNase H activity: No Fragment size: ~ 12.5 kb Application First-strand synthesis of cdna from RNA molecules (RT) RT-PCR Random priming reactions cdna library construction Probe labeling mrna 5 -end mapping by primer extension analysis Real-Time PCR Storage Temperature -20 Experimental Data Figure 1. Performance of the AccuPower RocketScript TM RT PreMix, RNase H Minus in two-step RT-PCR. RT was used to amplify 500 bp to 11 kb cdna. Sizes are indicated above gel. The larger amplicons require 1 µg of RNA, likely due to the low abundance of full-length RNA and the difficulty in reading through a long transcript. 58

AccuPower RocketScript RT PreMix, RNase H Minus NEW Figure 2. High synthesis rate at 50 using AccuPower RocketScript TM RT PreMix, RNase H Minus. Synthesis of cdna at 50 or 42 for 10, 20, 30 and 60 min using 1 µg of 9 kb RNA transcript as a template using Bioneer cdna synthesis kit and other kits for first strand cdna synthesis. Reaction products were resolved on a 1% alkaline agarose gel. Bioneer cdna synthesis kit completed synthesis of 9 kb transcript in 10 min. Lane 1: RT 10 min Lane 2: RT 20 min Lane 3: RT 30 min Lane 4: RT 60 min Figure 4. Sensitivity comparison of the various target. Rxn. conditions: Conventional 30 min incubation at 50 deactivation at 95 for 5 min Lane 1: 10 ng of total RNA from HeLa cells Lane 2: 1 ng of total RNA from HeLa cells Lane 3: 100 pg of total RNA from HeLa cells Lane 4: 10 pg of total RNA from HeLa cells Figure 3. Thermal stability using AccuPower RocketScript TM RT PreMix, RNase H Minus. Synthesis of cdna at 50-70 for 30 mins using 100 ng, 10 ng, 1 ng and 100 pg of 600 bp RNA transcript as a template using RocketScript TM RNase H(-) RT PreMix. RocketScript TM RNase H(-) RT PreMix completed synthesis of 70. Ordering Information Cat. no. Product Description K-2221 AccuPower RocketScript TM RT PreMix, RNase H Minus, 0.2 ml thin-wall 8-tube strips with attached cap, 96 tubes, 20 µl/rxn K-2222 AccuPower RocketScript TM RT PreMix, RNase H Minus, 0.2 ml thin-wall 8-tube strips with attached cap, 480 tubes, 20 µl/rxn K-2223 AccuPower RocketScript TM RT PreMix, RNase H Minus, 0.2 ml thin-wall 8-tube strips with attached cap, 96 tubes, 50 µl/rxn K-2224 AccuPower RocketScript TM RT PreMix, RNase H Minus, 0.2 ml thin-wall 8-tube strips with attached cap, 480 tubes, 50 µl/rxn www.bioneer.co.kr 59

AccuPower RocketScript RT-PCR PreMix, RNase H Minus NEW RNase H activity 를제거하여 Long RNA 의 One-step RT-PCR Description AccuPower RocketScript TM RT-PCR PreMix, RNase H Minus 는 70 에서도반응이가능한 RocketScript TM RTase, RNase H Minus 와 ProFi Taq DNA polymerase, dntp, reaction buffer 등을진공건조시킨 one-step RT-PCR 제품입니다. RNase H activity 는 cdna 합성시에 cdna 와 hybrid 된주형의 RNA 를제거하는역할을하는데너무강력한 RNase H 의활성을갖게되면 cdna 합성신장부위에도영향을주기때문에긴 size 의 cdna 를합성할수없게됩니다. cdna 합성과 PCR 의 2 단계반응을하나의 tube 에서 one-step 으로수행하는제품입니다. RocketScript TM RTase, RNase H Minus 이용하여고온에서 RT 반응을수행하기때문에, 안정된 2 차구조의 template RNA 에서도효과적으로 RT-PCR product 를얻을수있습니다. Features and Benefits Elimination RNase H activity: RocketScript TM RTase, RNase H Minus 는 cdna 합성과정에서생성되는 RNA/DNA hybrid 를분해하는 RNase H 활성이없어긴 target 의 cdna 합성에적합하고높은민감도를가지므로낮은농도의 RNA 에서도향상된성능으로증폭할수있습니다. Thermostable Activity: 70 에서도활성을갖는뛰어난열안정성의 RocketScript TM RTase 적용으로 2 차구조 RNA 의 cdna 합성에적합합니다. cdna 합성온도는 42~70 까지다양한온도로설정할수있어효과적으로 cdna 를합성할수있습니다. Sensitivity: Cell, tissue 등에서추출한 human total RNA 의 10 pg 에서도 one-step RT-PCR (Target: ß-Actin) 이가능합니다. Long kb RT-PCR: Human total RNA 로 full-length size 가 9 kb (target: Human fib) target 도 one-step RT-PCR 이가능합니다. Specifications Enzyme: RocketScript TM RTase (RNase H-), ProFi Taq DNA Polymerase 5 to 3 exonuclease: Yes 3 to 5 exonuclease: Yes 3 - A Overhang: Yes RNase H Activity: No Fragment size: ~ 9 kb Application Low copies detection RNA virus detection Gene expression analysis Storage Temperature -20 Ordering Information Cat. no. K-2231 K-2232 K-2233 K-2234 Product Description AccuPower RocketScript TM RT-PCR PreMix, RNase H Minus, 0.2 ml thin-wall 8-tube strips with attached cap/96 tubes, 20 μl/rxn AccuPower RocketScript TM RT-PCR PreMix, RNase H Minus, 0.2 ml thin-wall 8-tube strips with attached cap/480 tubes, 20 μl/rxn AccuPower RocketScript TM RT-PCR PreMix, RNase H Minus, 0.2 ml thin-wall 8-tube strips with attached cap/96 tubes, 50 μl/rxn AccuPower RocketScript TM RT-PCR PreMix, RNase H Minus, 0.2 ml thin-wall 8-tube strips with attached cap/480 tubes, 50 μl/rxn Ease-of-use: 각각의 PCR tube 에 RocketScript TM RTase, RNase H Minus, ProFi Taq DNA polymerase 및 RNase inhibitor 등 cdna 합성과 PCR 에필요한모든구성성분이포함되어있어 template RNA, primer set, D.W. 만을넣어바로 one-step RT-PCR 반응을수행할수있습니다. 또한전기영동시에필요한 tracking dye 와침강제가포함되어있어 sample loading buffer 를추가로넣을필요가없으므로편리하게사용할수있습니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. 60

AccuPower RocketPlex RT-PCR PreMix 한튜브에서 10 개의 Target 을동시검출하는 Multiplex One-step RT-PCR Specificity: HotStart Top DNA polymerase 를이용하여 PCR 반응의특이성을높였습니다. Description AccuPower RocketPlex RT-PCR PreMix 는하나의 tube 내에서두개이상의 target gene 의 cdna 합성과 PCR 을연속적으로수행할수있도록고안된제품입니다. 최대 10 개 target RNA 를동시에 one-step RT-PCR 할수있습니다. 본제품은바이오니아에서개발한 RocketScript TM Reverse Transcriptase 를사용한제품으로기존 M-MLV 와같은 RTase 가가지고있는열안정성문제를해결해줌으로써안정된 2 차구조의 RNA 를고온에서반응시켜효과적으로 full-length cdna 를합성할수있습니다. 또한, 각 tube 에 1 회사용분량이분주된 single use 제품으로, 반복사용을하는 master mix 제품에서발생하는 cross contamination 문제를방지합니다. Ease-of-use: 각각의 PCR tube에 RocketScript TM RTase, Top DNA polymerase 및 cdna 합성과 PCR에필요한모든구성성분이포함되어있어 template RNA, primers, D.W. 만을넣어바로 onestep multiplex RT-PCR 반응을수행할수있습니다. 또한전기영동시에필요한 tracking dye와침강제가포함되어있어 sample loading buffer를첨가할필요가없으므로간편하게사용할수있습니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system으로대량생산되어각 batch에대한철저한 QC를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. Specifications Enzyme: RocketScript TM RTase, HotStart Top DNA polymerase 5 to 3 exonuclease: No 3 to 5 exonuclease: No 3 - A overhang: Yes RNase H activity: Yes Fragment size: ~ 1 kb Application Multiplex RT-PCR Low copy detection Gene expression analysis Note: Schematic representation of the 5 UTR of a gene, with complex secondary structure, at three different temperatures. Note that RocketScript TM RTase shows full activity at 70 allowing it to synthesize the complete gene sequence where M-MLV and other reverse transcriptases fail. Storage Temperature -20 Experimental Data Features and Benefits Multiplex Performance: 하나의 tube 내에서 1~10 개의서로다른 target gene 에대하여 cdna 합성과증폭이가능합니다. Thermostable Activity: 70 에서도활성을갖는뛰어난열안정성의 RocketScript TM RTase 적용으로 2 차구조 RNA 의 cdna 합성에적합합니다. M-MLV RTase 효소의경우낮은열안정성을가지며, 이에따라역전사반응은낮은온도 (42 ) 에서만진행됨으로써안정된 2 차구조 RNA 의 cdna 합성효율이떨어지는단점이있습니다. 이러한문제점을해결하기위해 50 이상의높은온도에서도활성을갖는 reverse transcriptase 를자체개발하였습니다. 고객의실험조건에맞게 42~70 까지다양한온도에서 RT 반응을수행할수있어효과적으로 cdna 를합성할수있습니다. Figure 1. Single RT-PCR and multiplex RT-PCR using AccuPower RocketPlex RT-PCR PreMix. M: 25/100 bp Mixed DNA Ladder (Bioneer, Cat. no. D-1020) www.bioneer.co.kr 61

AccuPower RocketPlex RT-PCR PreMix Figure 2. Comparison of amplification quality between AccuPower RocketPlex RT-PCR PreMix and other suppliers RT-PCR kit. 10-plex primers were added into AccuPower RocketPlex RT-PCR PreMix and other supplier s RT-PCR kit. A series of human total RNA diluents were tested. All data were obtained using MyGenie TM 96 Gradient Thermal Block (Bioneer, Cat. no. A-2040-1). AccuPower RocketPlex RT-PCR PreMix is able to perform reverse transcription reactions throughout a wide range of temperatures from 42 to 70 Lane 1: Human total RNA 100 ng Lane 2: Human total RNA 10 ng Lane 3: Human total RNA 1 ng. 62

AccuPower RocketPlex RT-PCR PreMix Ordering Information Cat. no. Product Description K-2211 AccuPower RocketPlex RT-PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap/96 tubes, 20 μl/rxn K-2212 AccuPower RocketPlex RT-PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap/480 tubes, 20 μl/rxn K-2213 AccuPower RocketPlex RT-PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap/96 tubes, 50 μl/rxn K-2214 AccuPower RocketPlex RT-PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap/480 tubes, 50 μl/rxn www.bioneer.co.kr 63

AccuPower Dual-HotStart TM RT-PCR PreMix RT 와 PCR 에 HotStart 를적용하여높은특이성 / 민감도로 2 차구조 RNA 의 One-step RT-PCR Specifications Enzyme: RocketScript TM RTase, HotStart Taq DNA polymerase 5 to 3 exonuclease: Yes 3 to 5 exonuclease: No 3 - A overhang: Yes RNase H activity: Yes Fragment size: ~ 3 kb Description AccuPower Dual-HotStart TM RT-PCR PreMix 는종래에비특이적으로일어나는역전사반응의문제점들을근본적으로개선한 HotStart 역전사기술이적용된제품입니다. 원하는 RNA 만선택적으로역전사하므로민감도가획기적으로개선되어 template RNA 가극미량인시료에효과적입니다. 또한선택적으로역전사된 cdna 에 HotStart PCR 반응을수행하여원하는 Target 유전자를고감도로정확하게검출할수있는 one-step RT-PCR 제품입니다. 고감도 one-step RT-PCR 제품을이용하여 RNA 를대상으로하는각종바이러스검사및 gene expression 분석실험등에유용하게사용할수있습니다. 각 tube 에 1 회사용분량이분주된 single use 제품으로, 반복사용을하는 master mix 제품에서발생하는 cross contamination 문제를방지합니다. Application Low copy viral/bacterial pathogen load determination in an earlier stage Low copy mrna amplification Low copy target RNA quantification RNA amplification for microarray and NGS Storage Temperature -20 Accurate results! Excellent reproducibility! World-class technology! - HotStartRT + HotStart PCR = Dual-HotStart RT-PCR HotStart RT reaction HotStart PCR reaction Features and Benefits Specificity: PyroHot Start RT 반응과 HotStart PCR 을이용한세계최초 Dual-HotStart RT-PCR 반응은 target 유전자만을정확하게검출할수있도록최적화된제품입니다 (Figure 1). Sensitivity: 일반적으로검출이어려운극미량의 target template 가고농도의 RNA 에포함되어있는시료에서도검출이가능하며, 반응성이우수하여다양한종류의 PCR inhibitor 를포함하고있는혈액, 토양시료의 template RNA 에서도정확한 RT-PCR 결과를얻을수있습니다. Figure 1. Dual-HotStart TM technology. Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. Ease-of-use: 각각의 PCR tube 에 RocketScript TM RTase, HotStart Taq DNA polymerase 및 cdna 합성과 PCR 에필요한모든구성성분이포함되어있어 template RNA, primer set, D.W. 만을넣어바로 one-step RT-PCR 반응을수행할수있습니다. 또한전기영동시에필요한 tracking dye 와침강제가포함되어있어 sample loading buffer 를첨가할필요가없으므로간편하게사용할수있습니다. 64

AccuPower Dual-HotStart TM RT-PCR PreMix Experimental Data Figure 2. Comparison of PCR amplification sensitivity between AccuPower Dual-HotStart TM RT-PCR PreMix from Bioneer and other suppliers Hotstart RT-PCR kit. Target: human PGK1 Lane 1: 10 ng of human RNA Lane 2: 1 ng of human RNA Lane 3: 100 pg of human RNA Lane 4: 10 pg of human RNA Ordering Information Cat. no. Product Description K-6710 AccuPower Dual-HotStart TM RT-PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap/96 tubes, 20 ul/rxn K-6711 AccuPower Dual-HotStart TM RT-PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap/96 tubes, 50 ul/rxn K-6712 AccuPower Dual-HotStart TM RT-PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap/480 tubes, 20 ul/rxn K-6713 AccuPower Dual-HotStart TM RT-PCR PreMix, 0.2 ml thin-wall 8-tube strips with attached cap/480 tubes, 50 ul/rxn www.bioneer.co.kr 65

AccuPower Dual-HotStart TM RT-PCR PreMix (with UDG) Carryover Contamination 을제거. 높은특이성 / 민감도로 2 차구조 RNA 의 One-step RT-PCR Ease-of-use: 각각의 PCR tube 에 RocketScript TM RTase, HotStart Taq DNA polymerase 및 cdna 합성과 PCR 에필요한모든구성성분이포함되어있어 template RNA, primer set, D.W. 만을넣어바로 one-step RT-PCR 반응을수행할수있습니다. 또한전기영동시에필요한 tracking dye 와침강제가포함되어있어 sample loading buffer 를첨가할필요가없으므로간편하게사용할수있습니다. Description AccuPower Dual-HotStart TM RT-PCR PreMix (with UDG) 는종래에비특이적으로일어나는역전사반응의문제점들을근본적으로개선한 HotStart 역전사기술과 PCR 실험시야기될수있는 carryover contamination 문제를해결하는 uracil DNA glycosylase system 을함께적용한제품입니다. 본제품은 primerdimer 형성을최소화하여반응특이성과 PCR 증폭효율을높임과동시에위양성문제를해결하여각종바이러스검사및 gene expression 분석실험에유용하게사용할수있는고감도 onestep RT-PCR 제품입니다. 또한, 각 tube 에 1 회사용분량이분주된 single use 제품으로, 반복사용을하는 master mix 제품에서발생하는 cross contamination 문제를방지합니다. Specifications Enzyme: RocketScript TM RTase, HotStart Taq DNA polymerase 5 to 3 exonuclease: Yes 3 to 5 exonuclease: No 3 - A overhang: Yes RNase H activity: Yes Fragment size: ~ 3 kb Application Low copy viral/bacterial pathogen load determination in an earlier stage Low copy mrna amplification Low copy target RNA quantification RNA amplification for microarray and NGS Storage Temperature -20 Features and Benefits Prevention of Carryover Contamination: Uracil DNA Glycosylase (UDG) 는 DNA 가닥내 uracil 과 deoxyribose 간의 N-glycosylic bond 를가수분해함으로써 uracil 이삽입된 template 을제거합니다. UDG 는 PCR cycling 전 37 에서 2 분반응을통해활성화되어, uracil 이들어간 DNA 를제거하므로공기중남아있던산물의유입에의한 carryover 오염을방지할수있으며, PCR cycling 중고온에서불활성화됩니다 (Figure 1). Specificity: PyroHotStart RT 반응과 HotStart PCR 을이용한세계최초 Dual-HotStart RT-PCR 반응은 target 유전자만을정확하게검출할수있도록최적화된제품입니다 (Figure 2). Sensitivity: 일반적으로검출이어려운극미량의 target template 가고농도의 RNA 에포함되어있는시료에서도검출이가능하며, 반응성이우수하여다양한종류의 PCR inhibitor 를포함하고있는혈액, 토양시료의 template RNA 에서도정확한 RT-PCR 결과를얻을수있습니다. Figure 1. Prevention of carryover contamination. Accurate results! Excellent reproducibility! World-class technology! - HotStartRT + HotStart PCR = Dual-HotStart RT-PCR HotStart RT reaction HotStart PCR reaction Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. Figure 2. Dual-HotStart technology 66

AccuPower Dual-HotStart TM RT-PCR PreMix (with UDG) Experimental Data RNA template (not including uracil base) RNA template (including uracil base) Figure 4. Comparison of PCR amplification specificity between AccuPower Dual-HotStart TM RT- PCR PreMix (with UDG) from Bioneer and other suppliers Hotstart PCR master mix. Lane 1: HCV 10 7 Lane 2: HCV 10 6 Lane 3: HCV 10 5 Lane 4: HCV 10 4 Figure 3. Comparison of amplification quality using PCR products (not including uracil base or including uracil base) between AccuPower Dual-HotStart TM RT-PCR PreMix and AccuPower Dual-HotStart TM RT- PCR PreMix (with UDG). Lane 1: 10 8 Lane 2: 10 7 Lane 3: 10 6 Lane 4: 10 5 Lane 5: 10 4 Lane 6: 10 3 Lane 7: 10 2 Lane N: NTC Ordering Information Cat. no. Product Description K-6714 AccuPower Dual-HotStart TM RT-PCR PreMix (with UDG), 0.2 ml thin-wall 8-tube strips with attached cap/96 tubes, 20 ul/rxn K-6715 AccuPower Dual-HotStart TM RT-PCR PreMix (with UDG), 0.2 ml thin-wall 8-tube strips with attached cap/96 tubes, 50 ul/rxn www.bioneer.co.kr 67

M-MLV Reverse Transcriptase Standard cdna Synthesis Application First strand cdna synthesis from RNA, RT-PCR, and qrt-pcr Reagents Supplied 5 x Reaction Buffer (ph 8.1): Tris-HCl, KCl, etc. 100 mm DTT dntps mixture: 10 mm, each dntp 2.5 mm Concentration 10,000 U (200 U/μl) Description Moloney Murine Leukemia Virus (M-MLV) 에서유래된 RNA dependent DNA polymerase 로써 RNA 를주형으로 first strand cdna 를합성할수있습니다. cdna 를합성한후 M-MLV reverse transcriptase 의 RNase H+ 활성에의해 template RNA 가제거되므로, 잔존하는 template RNA 에의한영향을최소화할수있습니다. Features and Benefits Optimized 5X buffer 제공 : cdna 합성이 10분이내에가능합니다. Full length cdna 합성 : 최대 9 kb까지합성이가능합니다. Reproducibility: 재현성있는결과를위해바이오니아의전제품은엄격한 ISO 품질시스템하에서생산됩니다. Storage Conditions ph 7.6, Tris-Cl, NaCl, EDTA, DTT, Stabilizers, etc. Storage Temperature -20 Unit Definition One unit is defined as the amount of enzyme required to incorporate 1 nmole of dttp into acid-precipitable material in 10 min at 37 using poly A, oligo dt as template primer. Specifications DNase activity : No RNase acitivity: No RNase H activity: Yes Strand displacement: Yes Fragment size: ~9 kb Ordering Information Cat. no. Product Description E-3121 M-MLV Reverse Transcriptase, 10,000 U, 5 x Reaction Buffer, 100 mm DTT, 10 mm dntps E-3122 M-MLV Reverse Transcriptase, 50,000 U, 5 x Reaction Buffer, 100 mm DTT, 10 mm dntps 68

CycleScript Reverse Transcriptase High Performance cdna Synthesis Description CycleScript Reverse Transcriptase (CTase) 는고온에서도 cdna 를합성할수있는제품으로고순도로정제된역전사효소에안정화물질을첨가하였습니다. 본제품은기존의 42 단일온도역전사반응 (Fixed Temperature Reverse Transcription, FTRT) 뿐만아니라, PCR 반응과같이 2-3 단계의순차적인온도변화를통한순환온도역전사반응 (Cyclic Temperature Reverse Transcription, CTRT) 을수행할수있는제품입니다. 단일온도역전사반응 (FTRT) 및순환적역전사반응 (CTRT) 과정은다음과같습니다. 상기단일온도역전사반응 (FTRT) 및순환적역전사반응 (CTRT) 과정은다음과같습니다. 단일온도역전사반응 (FTRT) - Step 1: RNA denaturation 단계 (65, 10 min) - Step 2: cdna 합성단계 (42, 15~60 min) 순환온도역전사반응 (CTRT) - Step 1: Primer annealing 단계 (25~40, 30 sec) - Step 2: cdna 합성단계 (42~48, 4 min) - Step 3 (optional): RNA template 2차구조해소및 cdna 합성단계 (50~55, 30 sec) * 상기반응에서온도, 시간및 cycle 은고객의실험조건에따라선택적으로설정할수있습니다. 바이오니아가특허출원한순환온도역전사반응은저온반응 (15~40 ) 에서 primer annealing을유도하고, 고온반응 (50~55 ) 에서 template RNA 이차구조형성을해소함으로써기존의 42 역전사반응보다효율적으로 cdna를합성할수있습니다. 또한, 역전사반응에필요한 DTT, CycleScript Reverse Transcriptase, reaction buffer외에 dntps mixture도포함하고있어별도의구매가필요하지않습니다. Features and Benefits Stability: RNase-, DNase- & Proteinase-free의순수한 reverse transcriptase (RTase, M-MLV) 에안정화물질을첨가하여효소의열안정성을높였습니다. High Efficiency: 저온반응에서 primer annealing 을유도하고, 고온에서 RNA template 의 2 차구조를해소하여 cdna 합성율의증가시키고 full length cdna 합성을가능하게합니다. Flexible Reaction Conditions: CycleScript Reverse Transcriptase 는열안정성을높인제품으로, 단일온도역전사반응뿐만아니라순환온도역전사반응에적용할수있습니다. Ease-of-use & High Speed: 순환적역전사반응을수행할경우 primer 와 RNA template 의 pre-incubation 과정이생략되어실험방법이간단하고반응시간이단축됩니다. Reproducibility: 재현성있는결과를위해바이오니아의전제품은엄격한 ISO 품질시스템하에서생산됩니다. Specifications DNase activity : No RNase acitivity: No RNase H activity: Yes Strand displacement: Yes Fragment size: ~9 kb www.bioneer.co.kr 69

CycleScript Reverse Transcriptase Application First-strand synthesis of cdna from RNA molecules RT-PCR Random priming reaction Library construction Probe labeling mrna 5 end mapping by primer extension analysis Reagents Supplied 5 x Reaction Buffer (ph 8.1): Tris-HCl, KCl, etc. 100 mm DTT dntps mixture: 10 mm, each dntp 2.5 mm Storage Conditions ph 7.6, Tris-Cl, NaCl, EDTA, DTT, Stabilizers, etc. Storage Temperature -20 Unit Definition One unit is defined as the amount of enzyme required to incorporate 1 nmole of dttp into acid-precipitable material in 10 min at 37 using poly A oligo dt as template primer. Concentration 10,000 U (200 U/µl) Principles 일반적으로사용되는 oligo dt 및 random primer 는 Tm 값이 15~40 정도로낮아기존의 42 역전사반응에서는 template RNA 와충분히 annealing 하기어렵고, cdna 합성에장애가되는 template RNA 2 차구조가유지되는경향이있습니다. 바이오니아의특허기술인순환온도역전사반응은저온반응 (15~40 ) 에서 primer annealing 을유도하고고온반응 (50~55 ) 에서 template RNA 2 차구조형성을해소함으로써효율적으로 cdna 를합성할수있습니다. 70

CycleScript Reverse Transcriptase Experimental Data Figure 1. Comparison of transferrin receptor gene amplification with different reverse transcriptases. 700 ng of total RNA was used for reverse transcription and the same amount of amplified products were used for electrophoresis. Lane 1-4: TFR (Transferrin receptor gene) amplified with M-MLV Lane 5-8: TFR amplified with CycleScript Lane 9-12: TFR amplified with CycleScript Lane 13-16: TFR amplified with M-MLV from Company I Lane 17-20: TFR amplified with S-script from Company S Lane 21-24: TFR amplified with S-script ll from Company I Lane 25-28: TFR amplified with S-script lll from Company I Lane 29-32: TFR amplified with O-script from Company Q Lane M.W.: 100 bp Plus DNA Ladder (Bioneer, Cat. no. D-1035) Figure 3. Working temperature comparison of different reverse transcriptases. Each 10 ng, 1 ng, 100 pg, and 10 pg of total RNA was used for reverse transcription and the same amount of amplified products were used for electrophoresis. Lane 1-4: GAPDH amplified with CycleScript Lane 5-8: GAPDH amplified with CycleScript Lane 9-12: GAPDH amplified with CycleScript Lane 13-16: GAPDH amplified with S-script from company S Lane 17-20: GAPDH amplified with S-script from company S Lane 21-24: GAPDH amplified with S-script from company S Figure 2. Comparison of GAPDH gene amplification with different reverse transcriptases. Each 10 ng, 1 ng, 100 pg, and 10 pg of total RNA was used for reverse transcription and the same amount of amplified products were used for electrophoresis. Lane 1-4: GAPDH amplified with CycleScript Lane 5-8: GAPDH amplified with CycleScript Lane 9-12: GAPDH amplified with CycleScript Lane 13-16: GAPDH amplified with M-MLV from Company I Lane 17-20: GAPDH amplified with S-script from Company S Lane 21-24: GAPDH amplified with S-script from Company S Lane 25-28: GAPDH amplified with S-script from Company I Lane 29-32: GAPDH amplified with S-script from Company I Ordering Information Cat. no. Product Description E-3131 CycleScript Reverse Transcriptase, 10,000 U, 10 mm dntps, 5 x Reaction Buffer, 100 mm DTT E-3132 CycleScript Reverse Transcriptase, 50,000 U, 10 mm dntps, 5 x Reaction Buffer, 100 mm DTT www.bioneer.co.kr 71

RocketScript Reverse Transcriptase High Performance / High Temperature cdna Synthesis Description Rocketscript TM Reverse Transcriptase 는바이오니아가독자적으로개발한 M-MLV 유래의 highly thermostable reverse transcriptase 입니다. 저온 (42 ) 에서최적활성을갖는 M-MLV RTase 는복잡한 2 차구조를가진 RNA 로부터 cdna 를합성하는데많은문제점이있습니다. Rocketscript TM RTase 는 thermostable activity (42~70 ) 를갖고있어복잡한 2 차구조의 RNA 로부터효과적으로 full-length cdna 를합성할수있습니다. Features and Benefits Thermostable Activity: 열안정성이낮은 M-MLV RTase 효소는낮은온도 (42 ) 에서만역전사반응이진행되기때문에복잡한고차구조의 RNA를 cdna로합성하는경우반응이제대로진행되지않는단점이있습니다. 이러한문제점을해결하기위해바이오니아는 synthetic biology 기술을이용하여 50 이상의높은온도에서도활성을갖는 reverse transcriptase를개발하였습니다. 기존 42 RT 반응에만국한되었던한계를극복한 Rocketscript TM series 제품은 42~70 까지고객의실험조건에맞게다양한온도에서 RT 반응을수행할수있어효과적으로 cdna를합성할수있습니다. Specifications DNase activity : No RNase acitivity: No RNase H activity: Yes Fragment size: ~10 kb Application Gene synthesis First-strand synthesis of cdna from RNA molecules (Reverse Transcription) RT-PCR Random priming reactions Library construction Probe labeling mrna 5 -end mapping by primer extension analysis Real-Time PCR Concentration 10,000 U (200 U/μl) Storage Temperature -20 Unit Definition One unit is defined as the amount of enzyme required to incorporate 1 nmole of dttp into acid-precipitable material in 10 min at 37 using poly A, oligo dt as template primer. Reproducibility: 재현성있는결과를위해바이오니아의전제품은엄격한 ISO 품질시스템하에서생산됩니다. 72

RocketScript Reverse Transcriptase Experimental Data Figure 2. Thermostable stability check. Amplification results of RocketScript TM Reverse Transcriptase using myc compared with supplier I Reverse transcription. Rxn. condition: Incubation at each temperature 45 /50 /55 /60 /65 /70 for 1 hr, deactivation at 95 for 5 min Target: human myc 495 bp Lane 1: 100 ng human total RNA from HeLa cell Lane 2: 10 ng human total RNA from HeLa cell Lane 3: 1 ng human total RNA from HeLa cell Lane 4: 100 pg human total RNA from HeLa cell Lane M: 1 kb DNA Ladder (Bioneer, Cat. no. D-1040) Figure 1. Complex RNA amplification. Complex RNA amplification results of RocketScript TM Reverse Transcriptase compared with that of conventional RTase. Rxn. condition: conventional 42 / 50 / 60 1 hr, deactivation 95 5 min, this product shows thermal stability. Lane A: M-MLV Reverse Transcriptase Lane B: RocketScript TM Reverse Transcriptase Lane 1: 100 ng human total RNA from HeLa cell Lane 2: 10 ng human total RNA from HeLa cell Figure 3. Comparison of amplification quality between RocketScript TM RTase and competitor RTases. Target gene expression Level. Lane 1: 100 ng human total RNA from HeLa cell Lane 2: 10 ng human total RNA from HeLa cell Lane 3: 1 ng human total RNA from HeLa cell Lane 4: 100 pg human total RNA from HeLa cell Lane M: 1 kb DNA Ladder (Bioneer, Cat. no. D-1040) Ordering Information Cat. no. Product Description E-3141 RocketScript TM Reverse Transcriptase, 10,000 U (50 rxns) E-3142 RocketScript TM Reverse Transcriptase, 50,000 U (250 rxns) www.bioneer.co.kr 73

RocketScript Reverse Transcriptase, RNase H Minus NEW High Performance / High Temperature / Long Size cdna Synthesis Specifications DNase activity : No RNase acitivity: No RNase H activity: No Fragment size: ~12.5 kb Description Rocketscript TM Reverse Transcriptase, RNase H Minus 는바이오니아가독자적으로개발한 Rocketscript TM Reverse Trancripatase 를 mutation 시켜 RNase H 의활성을제거한제품입니다. RNase H 활성을갖게되면 cdna 합성신장부위에영향을주기때문에긴 size 의 cdna 를합성할수없습니다. Rocketscript TM Reverse Transcriptase, RNase H Minus 는 RNase H 활성이없기때문에긴 size 의 cdna 를합성할수있습니다. Features and Benefits RNase H Activity: Rocketscript TM series 제품은 42~70 까지고객의실험조건에맞게다양한온도에서 RT 반응을수행할수있어효과적으로 cdna를합성할수있지만, RNase H 활성이있어긴 size의 cdna를합성할수없습니다. 본제품은 point mutation을통해 RTase H의활성을제거하여 ~12.5 kb까지효과적으로 cdna를합성할수있습니다. Thermostable Activity: Rocketscript TM Reverse Transcriptase 를기반으로개발한제품으로써 42~70 까지고객의실험조건에맞게다양한온도에서 RT 반응을수행할수있어효과적으로 cdna 를합성할수있습니다. Reproducibility: 재현성있는결과를위해바이오니아의전제품은엄격한 ISO 품질시스템하에서생산됩니다. Application Gene synthesis First-strand synthesis of cdna from RNA molecules (Reverse Transcription) RT-PCR Random priming reactions Library construction Probe labeling mrna 5 -end mapping by primer extension analysis Real-Time PCR Concentration 10,000 U (200 U/μl) Storage Temperature -20 Unit Definition One unit is defined as the amount of enzyme required to incorporate 1 nmole of dttp into acid-precipitable material in 10 min at 37 using poly A, oligo dt as template primer. 74

RocketScript Reverse Transcriptase, RNase H Minus NEW Experimental Data st strand cdna synthesis kit in two-step RT-PCR. RT was used to amplify 500 bp to 11 kb cdna. Sizes are indicated above gel. The larger amplicons require 1 μg of RNA, likely due to the low abundance of full-length RNA and the difficulty in reading through a long transcript Ordering Information Figure 2. High synthesis rate at 50 using Bioneer first strand cdna synthesis kit. Synthesis of cdna at 50 or 42 for 10, 20, 30 and 60 minutes using 1 μg of 9 kb RNA transcript as a template using Bioneer cdna Synthesis Kit and other kits for first strand cdna synthesis. Reaction products were resolved on a 1% alkaline agarose gel. Bioneer cdna synthesis kit completed synthesis of 9 kb transcript in 10 min. Lane 1: RT 10 min Lane 2: RT 20 min Lane 3: RT 30 min Lane 4: RT 60 min Cat. no. Product Description E-3151 RocketScript TM Reverse Transcriptase, RNase H Minus, 10,000 U (50 rxns) E-3152 RocketScript TM Reverse Transcriptase RNase H Minus, 50,000 U (250 rxns) www.bioneer.co.kr 75

03 Real-Time PCR Intercalating Dye Type Kit AccuPower GreenStar qpcr PreMix 77 AccuPower 2X GreenStar qpcr Master Mix 79 TaqMan Probe Type Kit AccuPower DualStar qpcr PreMix 81 AccuPower Plus DualStar qpcr PreMix & 2X Master Mix 83 AccuPower Plus DualStar qpcr PreMix & 2X Master Mix (with UDG) New 85 AccuPower Dual-HotStart RT-qPCR PreMix & 2X Master Mix 87 Real-Time PCR Instrument Exicycler Go to M. Instruments & Devices

AccuPower GreenStar qpcr PreMix PyroHotStart 기술을적용한 Intercalating Dye 방식의 Real-Time PCR Application Real-Time quantification of DNA and cdna targets Gene expression profiling Gene functional analysis Microbial & viral pathogen detection Description AccuPower GreenStar TM qpcr PreMix는 intercalating dye 와바이오니아의 enzyme-mediated HotStart 특허기술을적용하여반응특이성과 PCR 증폭효율을높인 Real-Time PCR 제품입니다. 또한 Real-Time PCR에필요한모든시약을반응 tube에진공건조시킨제품으로, 반복사용을하는 master mix 제품에서발생하는 cross contamination 문제를방지하고, 간편하고정확한실험결과를얻을수있습니다. Features and Benefits Specificity: 바이오니아의특허기술 (enzyme-mediated HotStart 방법 ) 을적용하여반응전 (zero cycle) 의비특이증폭산물생성을최소화하고, 매 cycle 마다생성되는 PCR inhibitor (PPi) 를가수분해하여 PCR 반응효율을극대화함으로써미량의 template DNA 를효과적으로증폭할수있습니다 (Figure 1). Figure 1. Enzyme-mediated HotStart PCR. Experimental Data High Specificity Ease-of-use: 각각의 PCR tube에 DNA polymerase와 Real-Time PCR 수행에필요한모든구성성분이포함되어있어 template DNA, primer set 및 D.W. 만넣어바로 Real-Time PCR 반응을수행할수있습니다. Stability: PCR 반응혼합물에안정화제를첨가하여진공건조시켜, -20 에서 2 년동안장기보관하여도활성을안정적으로유지합니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. Specifications Enzyme: Top DNA polymerase 5 to 3 exonuclease: No 3 to 5 exonuclease: No 3 - A overhang: Yes Figure 2. Real-time PCR data of AccuPower GreenStar TM qpcr PreMix. AccuPower GreenStar TM qpcr PreMix provides at least 7 orders of magnitude in dynamic range (10 fg ~ 10 ng /rxn). A: Amplification curve of AccuPower GreenStar TM qpcr PreMix. Lambda DNA primers were added into AccuPower GreenStar TM qpcr PreMix. A series of lambda DNA positive control diluents were tested. B: Standard curve of AccuPower GreenStar TM qpcr PreMix. R 2-0.998, PCR efficiency - 95% C: Melting curve analysis of AccuPower GreenStar TM qpcr PreMix. The melting curve shows that only single amplified PCR product was obtained in all template range. All data were obtained using Exicycler TM 96 Real-Time Quantitative Thermal Block (Bioneer, Cat. no. A-2060). www.bioneer.co.kr 77

AccuPower GreenStar qpcr PreMix Procedure General reaction condition Components Concentration 20 μl rxn 50 μl rxn Template 10 9 ~ 10 3 copies/rxn 5 μl 5 μl F-primer (10 pmole/μl) Each 0.25 ~ 1 μm 1 ~ 2 μl 1 ~ 2 μl R-primer (10 pmole/μl) Each 0.25 ~ 1 μm 1 ~ 2 μl 1 ~ 2 μl DEPC-distilled water Adjust to 20 μl Adjust to 50 μl Figure 3. Comparison of amplification efficiency between Accu- Power GreenStar TM qpcr PreMix and other company s master mixture. Amplification curve of AccuPower GreenStar TM qpcr PreMix and other company s master mix kit. Lambda DNA primers were added into AccuPower GreenStar TM qpcr PreMix and other company s master mix kit. A series of Lambda DNA positive control diluents were tested. Reaction mixtures were prepared and qpcr was performed according to each company s protocol. All data were obtained using ABI7500 Fast Real-Time PCR machine (Applied Biosystems co.). Concentration of template DNA and primer Template DNA and Conventional PCR primer Lambda DNA: 1 pg ~ 100 ng Template Bacteria DNA: 100 pg ~ 100 ng Human DNA: 1 ng ~ 500 ng Primer 5 ~ 30 pmoles Ordering Information Cat. no. Product Description K-6200 AccuPower GreenStar TM qpcr PreMix, 50 μl/rxn, 8-tube strips, 96 rxn, Exicycler TM 96, optical film included K-6201 AccuPower GreenStar TM qpcr PreMix, 50 μl/rxn, 8-tube strips, 96 rxn, ABI7500, optical film included K-6202 AccuPower GreenStar TM qpcr PreMix, 50 μl/rxn, 8-tube strips with cap, 96 rxn, Opticon K-6203 AccuPower GreenStar TM qpcr PreMix, 50 μl/rxn, 96-well plate, 96 rxn, Exicycler TM 96, optical film included K-6204 AccuPower GreenStar TM qpcr PreMix, 50 μl/rxn, 96-well plate, 96 rxn, ABI7500, optical film included K-6210 AccuPower GreenStar TM qpcr PreMix, 20 μl/rxn, 8-tube strips, 96 rxn, Exicycler TM 96, optical film included K-6211 AccuPower GreenStar TM qpcr PreMix, 20 μl/rxn, 8-tube strips, 96 rxn, ABI7500, optical film included K-6212 AccuPower GreenStar TM qpcr PreMix, 20 μl/rxn, 8-tube strips with cap, 96 rxn, Opticon K-6213 AccuPower GreenStar TM qpcr PreMix, 20 μl/rxn, 96-well plate, 96 rxn, Exicycler TM 96, optical film included K-6214 AccuPower GreenStar TM qpcr PreMix, 20 μl/rxn, 96-well plate, 96 rxn, ABI7500, optical film included 78

AccuPower 2X GreenStar qpcr Master Mix PyroHotStart 기술을적용한 Intercalating Dye 방식의 Real-Time PCR reaction chemicals 이 2X master mix 형태로포장되어손쉽게사용할수있습니다. 실험자는 2X master mix, template DNA, primers 및 D.W. 를혼합하여 Real-Time PCR 반응을수행할수있습니다. 50X Rox dye 가포함되어있어따로구매할필요가없습니다. Specifications Enzyme: Top DNA polymerase 5 to 3 exonuclease: No 3 to 5 exonuclease: No 3 - A overhang: Yes Description AccuPower 2X GreenStar TM qpcr Master Mix 는 intercalating dye, reaction buffer, dntps, 안정화제, DNA polymerase 와 hotstart PCR 반응에필요한모든성분을혼합한 2X master mix 형태로제공되는 Real-Time PCR 제품입니다. 바이오니아의독자적인 enzyme-mediated HotStart 특허기술을적용하여반응특이성과 PCR 증폭효율을높였습니다. 타사의 master mix 제품과는달리특허받은안정화제의첨가로본제품은 -20 에서 2 년까지효소활성이유지되어항상정확하고재현성있는 qpcr data 를얻을수있습니다. 또한 master mix type 으로여러 Real- Time PCR 장비에호환됩니다. Application Real-Time quantification of DNA and cdna targets Gene expression profiling Gene functional analysis Microbial & Viral pathogen detection Features and Benefits Specificity: 바이오니아의특허기술 (enzyme-mediated HotStart 방법 ) 을적용하여반응전 (zero cycle) 의비특이증폭산물생성및 primer dimer를최소화하고, 매 cycle 마다생성되는 PCR inhibitor (PPi) 를가수분해하여 PCR 반응효율을극대화함으로써미량의 template DNA를효과적으로증폭할수있습니다 (Figure 1). Figure 1. Enzyme-mediated HotStart PCR. Equipment Compatibility: Master mix type 의제품으로여러종류의 Real-Time PCR 장비에대해최적화된결과를얻을수있습니다. Dynamic Range: 10 ~ 10 8 copies 까지 8 log 의넓은 detection range 를갖습니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. Gene Expression Analysis: AccuPower 2X GreenStar TM qpcr Master Mix 는유전자발현양분석실험에매우적합한제품입니다. Ease-of-Use: Intercalating dye 를포함하여 PCR 반응에필요한모든 www.bioneer.co.kr 79

AccuPower 2X GreenStar qpcr Master Mix Experimental Data Equipment Compatibility Exicycler TM 96 of Target Gene hgapdh hptgs2 Sample No. 1 2 1 2 Ct Value 23.7. 23.46 33.98 30.52 Using a Bio-Rad IQ5 Figure 3. Gene expression analysis. AccuTarget TM Validated Real-Time PCR Primer Library for human is designed by Bioneer s bioinformatics tool and targeting for human genome. cdna was synthesized using human PTGS2 target primer of those and human total RNA identically quantified from Hela cell and blood cell with AccuPower CycleScript RT PreMix (Bioneer, Cat. no. K-2044). Gene analysis was carried out both Hela cell and blood cell by operating Real-Time PCR reaction (95 10 min, 1 cycle and 95 10 sec, 58 25 sec, 72 30 sec, 41 cycles) using the cdna, AccuPower 2X GreenStar TM PCR Master Mix and Exicycler TM 96 Real- Time Thermal Block (Bioneer, Cat. no. A-2060). Using a ABI7500 Figure 2. Comparison of the specificity of the intercalating dye based Real-Time PCR. Amplification of an 90 bp target gene was detected using serially diluted LP (Legionella Pneumoniae) genomic DNA (10 n dilution: 10 5 ~10 copies) with AccuPower 2X GreenStar TM qpcr Master Mix. As shown in figure, very small amount of primer dimers was appeared in AccuPower 2X GreenStar TM qpcr Master Mix than other kits. Ordering Information Cat. no. Product Description K-6251 AccuPower 2X GreenStar TM qpcr Master Mix, 50 μl/rxn, 100 rxn, 80X ROX Dye (0.1 ml X 1 ea) K-6252 AccuPower 2X GreenStar TM qpcr Master Mix, 50 μl/rxn, 200 rxn, 80X ROX Dye (0.1 ml X 1 ea) K-6253 AccuPower 2X GreenStar TM qpcr Master Mix, 50 μl/rxn, 100 rxn, without ROX Dye K-6254 AccuPower 2X GreenStar TM qpcr Master Mix, 50 μl/rxn, 200 rxn, without ROX Dye 80

AccuPower DualStar qpcr PreMix PyroHotStart 기술을적용한 Taqman Probe 방식의 Real-Time PCR Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. Specifications Enzyme: Taq DNA polymerase 5 to 3 exonuclease: Yes 3 to 5 exonuclease: No 3 - A overhang: Yes Description AccuPower DualStar TM PreMix 는 Taq DNA polymerase 의 5 ->3 exonuclease 활성을이용한 Taqman probe 기술이적용된 Real-Time PCR 제품입니다. 다양한종류의시료로부터고객이원하는 target 유전자를빠른시간내에정확하게정량할수있습니다. 바이오니아의 enzyme-mediated HotStart 특허기술을적용하여반응특이성과 PCR 증폭효율을높였습니다. Real- Time PCR 에필요한모든시약을반응 tube 에진공건조시킨제품으로, 반복사용을하는 master mix 제품에서발생하는 cross contamination 문제를방지하고, 간편하고정확한실험결과를얻을수있습니다. 또한, 냉동상태에서 2 년동안보관할수있으며, 여러종류의 Real-Time PCR 장비에서최상의결과를얻을수있도록다양한반응 tube 에건조하여제공합니다. Application Gene expression profiling Target DNA quantification Microbial detection Viral/bacterial pathogen load determination Evaluation of primer pair performance for probebased Real-Time PCR Features and Benefits Ease-of-use: 각각의 PCR tube 에 DNA polymerase 와 Real-Time PCR 수행에필요한모든구성성분이포함되어있어 template DNA, target 유전자에대한 probe & primer, D.W. 만넣어바로 Real-Time PCR 반응을수행할수있습니다. Specificity: 바이오니아의특허기술 (enzyme-mediated HotStart 방법 ) 을적용하여반응전 (zero cycle) 의비특이증폭산물생성및 primer dimer 를최소화하고, 매 cycle 마다생성되는 PCR inhibitor (PPi) 를가수분해하여 PCR 반응효율을극대화함으로써미량의 template DNA 를효과적으로증폭할수있습니다 (Figure 1). Figure 1. Enzyme-mediated HotStart PCR. Experimental Data Equipment Compatibility: 여러종류의 Real-Time PCR 장비에대해최적화된결과를얻을수있습니다. Universality of target gene: DNA, cdna 와 high GC template 등유전자의종류에상관없이유효한정량 PCR 결과를얻을수있습니다. Stability: PCR 반응혼합물에안정화제를첨가하여진공건조시켜, -20 에서 2 년동안장기보관하여도활성을안정적으로유지합니다. Figure 2. Real-Time PCR data of AccuPower DualStar TM qpcr PreMix. AccuPower DualStar TM qpcr PreMix provides at least 7 orders of magnitude in dynamic range (10 7 ~ 10 copies/rxn). A: Amplification curve. West Nile Virus (WNV) primers and TaqManbased probe were added into AccuPower DualStar TM qpcr PreMix. A series of WNV positive control diluents were tested. B: Standard curve. All data were obtained using Exicycler TM 96 Real-Time Quantitative Thermal Block (Bioneer, Cat. no. A-2060). www.bioneer.co.kr 81

AccuPower DualStar qpcr PreMix Figure 3. Data using various kinds of Real-Time PCR Instruments. AccuPower DualStar TM qpcr PreMix is applicable to most of commercially available Real-Time quantitative PCR instruments. West Nile Virus (WNV) primers and TaqMan-based probe were added into AccuPower DualStar TM qpcr PreMix. A series of WNV positive control diluents were tested. A: Amplification curve and standard curve using ABI7500 Fast Real-Time PCR machine (Applied Biosystems Co.). B: Amplification curve and standard curve using ABI7500 Real-Time PCR machine (Applied Biosystems Co.). C: Amplification curve and standard curve using Opticon Real-Time PCR machine (MJ Research, currently Bio-Rad Inc.). Figure 4. Comparison of detection sensitivity between AccuPower DualStar TM qpcr PreMix and other suppliers master mixture. West Nile Virus (WNV) primers and TaqMan-based probe were added into AccuPower DualStar TM qpcr PreMix and other suppliers master mixture. A series of WNV positive control diluents were tested. Reaction mixtures were prepared and qpcr was performed according to each supplier s protocol. All data were obtained using ABI7500 Fast Real-Time PCR machine (Applied Biosystems co.). Ordering Information Cat. no. Product Description K-6100 AccuPower DualStar TM qpcr PreMix, 20 μl/rxn, 8-tube strips, 96 rxn, Exicycler TM 96, optical film included K-6101 AccuPower DualStar TM qpcr PreMix, 20 μl/rxn, 8-tube strips, 96 rxn, ABI7500, optical film included K-6102 AccuPower DualStar TM qpcr PreMix, 20 μl/rxn, 8-tube strips with cap, 96 rxn, Opticon K-6103 AccuPower DualStar TM qpcr PreMix, 20 μl/rxn, 96-well plate, 96 rxn, Exicycler TM 96, optical film included K-6104 AccuPower DualStar TM qpcr PreMix, 20 μl/rxn, 96-well plate, 96 rxn, ABI7500, optical film included K-6110 AccuPower DualStar TM qpcr PreMix, 50 μl/rxn, 8-tube strips, 96 rxn Exicycler TM 96, optical film included K-6111 AccuPower DualStar TM qpcr PreMix, 50 μl/rxn, 8-tube strips, 96 rxn, ABI7500, optical film included K-6112 AccuPower DualStar TM qpcr PreMix, 50 μl/rxn, 8-tube strips with cap, 96 rxn, Opticon K-6113 AccuPower DualStar TM qpcr PreMix, 50 μl/rxn, 96-well plate, 96 rxn, Exicycler TM 96, optical film included K-6114 AccuPower DualStar TM qpcr PreMix, 50 μl/rxn, 96-well plate, 96 rxn, ABI7500, optical film included 82

AccuPower Plus DualStar qpcr PreMix & 2X Master Mix HotStart 기술을적용한 Taqman Probe 방식의 Real-Time PCR Specifications Enzyme: HotStart Taq DNA polymerase 5 to 3 exonuclease activity: Yes 3 to 5 exonuclease activity: No 3 - A Overhang: Yes Application Gene expression profiling Target DNA quantification Microbial detection Viral/bacterial pathogen load determination Evaluation of primer pair performance for probebased Real-Time PCR Description AccuPower Plus DualStar TM qpcr PreMix 는 Taq DNA polymerase 의 5 ->3 exonuclease 활성을이용한 Taqman probe 기술이적용된 Real-Time PCR 제품입니다. Real-Time PCR 에필요한모든시약을반응 tube 에진공건조시킨제품으로, 반복사용을하는 master mix 제품에서발생하는 cross contamination 문제를방지하고, 간편하고정확한실험결과를얻을수있습니다. 또한, -20 에서 2 년동안보관이가능하며, master mix 제품은여러종류의 Real-Time PCR instrument 에서최상의결과를얻을수있도록최적화되었습니다. Storage Temperature -20 Experimental Data Specificity Features and Benefits Ease-of-use: 각각의 PCR tube에 DNA polymerase와 Real-Time PCR 수행에필요한모든구성성분이포함되어있어 template DNA, target 유전자에대한 probe & primer, D.W. 만넣어바로 Real-Time PCR 반응을수행할수있습니다. 또한, master mix 제품은여러종류의 Real-Time PCR 장비에적용하여최적화된결과를얻을수있습니다. Dynamic Range: 10~10 8 copies까지 8 log의넓은 detection range 를갖습니다. Specificity: HotStart가적용된 DNA polymerase는반응전 (zero cycle) 의비특이증폭산물의생성을최소화합니다. PCR 온도가증가하면서활성화된 DNA polymerase는정확한 target만을빠르게증폭합니다. Universality of target gene: DNA, cdna와 high GC template 등유전자의종류에상관없이유효한정량 PCR 결과를얻을수있습니다. Stability: PCR반응혼합물에안정화제를첨가하여진공건조시켜, -20 에서 2년동안장기보관하여도활성을안정적으로유지합니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system으로대량생산되어각 batch에대한철저한 QC를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. Figure 1. High specificity of AccuPower Plus DualStar TM qpcr PreMix. Template concentration: 1x10 ~ 1x10 6 copies/rxn. Sensitivity Templete DNA copies Ct Value A Company B Company 10 3 32.6 34.72 31.28 10 4 29.18 31.31 27.75 10 5 25.66 27.61 24.44 10 6 22.43 24.48 21.99 10 7 19.22 20.98 19.99 Efficiency/linearity 99%/1 96%/1 100%/0.9999 Figure 2. Comparison of amplification quality between AccuPower Plus DualStar TM qpcr PreMix and other supplier s Real-Time qpcr Kit. www.bioneer.co.kr 83

AccuPower Plus DualStar qpcr PreMix & 2X Master Mix Multiplexing Day Template DNA of copies 1.00E+07 1.00E+06 1.00E+05 1.00E+04 1.00E+03 1.00E+02 1.00E+01 NTC FAM 19.47 22.74 25.97 28.58 31.07 32.87 34.35 UD TET 19.38 22.59 25.87 28.62 31.06 33.37 34.6 UD TEXAT_RED 18.47 21.48 24.23 26.78 28.68 30.15 31.49 UD CY5 20.38 22.56 25.43 27.71 29.71 31.38 32.45 UD Figure 3. Four-target multiplexing on the Exicycler TM 96 instrument using AccuPower Plus DualStar TM qpcr PreMix. Figure shows amplification of a 4-target multiplex assay. The dyes used were FAM, TET, Texas Red and CY5, respectively. The data demonstrate that over a dilution series of input template, the AccuPower Plus DualStar TM qpcr PreMix can successfully and reliably generate up to 4-target multiplex data on the Exicycler TM 96. Ordering Information Cat. no. Product Description K-6600 AccuPower Plus DualStar TM qpcr PreMix, 50 μl/rxn, 8-tube strips, 96 rxn, Exicycler TM 96, optical film included, K-6601 AccuPower Plus DualStar TM qpcr PreMix, 50 μl/rxn, 8-tube strips, 96 rxn, ABI7500, optical film included K-6602 AccuPower Plus DualStar TM qpcr PreMix, 50/rxn, 8-tube strips with cap, 96 rxn, Opticon, optical film included K-6603 AccuPower Plus DualStar TM qpcr Master Mix (2X), 2.5 ml, 100 rxn 84

AccuPower Plus DualStar qpcr PreMix & 2X Master Mix (with UDG) Carryover Contamination 을제거한 Taqman Probe 방식의 HotStart Real-Time PCR Universality of target gene: DNA, cdna 와 high GC template 등유전자의종류에상관없이유효한정량 PCR 결과를얻을수있습니다. Stability: PCR 반응혼합물에안정화제를첨가하여진공건조시켜, -20 에서 2 년동안장기보관하여도활성을안정적으로유지합니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. Description AccuPower Plus DualStar TM qpcr PreMix 는 Taq DNA polymerase 의 5 ->3 exonuclease 활성을이용한 Taqman probe 기술이적용된 Real-Time PCR 제품입니다. Hotstart Taq DNA polymerase 를이용하여원하는 target 만선택적으로증폭할수있으며, 극미량의 template DNA 에서도우수한민감도를가지고있는동시에 uracil DNA glycosylase system 적용으로 carryover contamination 문제를해결하였습니다. 각 tube 에 1 회사용분량이분주된 single use 제품으로써 master mix 제품의반복사용에따른 cross contamination 을방지할수있으며, 별도의혼합단계가필요없어간편하게 PCR 을수행할수있습니다. Features and Benefits Prevention of Carryover Contamination: Uracil DNA Glycosylase (UDG) 는 DNA 가닥내 uracil과 deoxyribose 간의 N-glycosylic bond를가수분해함으로써 uracil 이삽입된 template을제거합니다. UDG는 PCR cycling 전 37 에서 2분반응을통해활성화되어, uracil이들어간 DNA를제거하므로공기중남아있던산물의유입에의한 carryover 오염을방지할수있으며, PCR cycling 중고온에서불활성화됩니다 (Figure 1). Specifications Enzyme: HotStart Taq DNA polymerase 5 to 3 exonuclease activity: Yes 3 to 5 exonuclease activity: No 3 - A Overhang: Yes Application Gene expression profiling Target DNA quantification Microbial detection Viral/bacterial pathogen load determination Evaluation of primer pair performance for probebased Real-Time PCR Storage Temperature -20 Ease-of-use: 각각의 PCR tube 에 DNA polymerase 와 Real-Time PCR 수행에필요한모든구성성분이포함되어있어 template DNA, target 유전자에대한 probe & primer, D.W. 만넣어바로 Real-Time PCR 반응을수행할수있습니다. 또한, master mix 제품은여러종류의 Real-Time PCR 장비에적용하여최적화된결과를얻을수있습니다. Dynamic Range: 10~10 8 copies 까지 8 log 의넓은 detection range 를갖습니다. Specificity: HotStart 가적용된 DNA polymerase 는반응전 (zero cycle) 의비특이증폭산물의생성을최소화합니다. PCR 온도가증가하면서활성화된 DNA polymerase 는정확한 target 만을빠르게증폭합니다. Figure 1. Prevention of carryover contamination. www.bioneer.co.kr 85

AccuPower Plus DualStar qpcr PreMix & 2X Master Mix (with UDG) Experimental Data PCR product type Linearity Efficiency 10 8 10 7 10 6 10 5 10 4 10 3 10 2 NTC dntps 0.9988 98 12.81 16.41 19.72 22.84 26.37 29.76 33.48 UD dntps including dutps - - UD UD UD UD UD UD UD UD Figure 2. Efficiency of uracil DNA glycosylase using PCR products (including dn/du base). Ct Value Company Linearity Efficiency 10 7 10 6 10 5 10 4 10 3 10 2 NTC Bioneer 0.9999 98% 15.73 19.14 22.99 25.91 28.96 33.1 UD A company 0.9994 97% 16.86 19.98 23.55 27.26 30.05 34.61 UD Figure 3. Comparison of amplification quality between AccuPower Plus DualStar TM qpcr PreMix (with UDG) and other supplier s Real-time qpcr kit. Ordering Information Cat. no. Product Description K-6605 AccuPower Plus DualStar TM qpcr PreMix (with UDG), 50 μl/rxn, 8-tube strips, 96 rxn, Exicycler TM 96, optical film included, K-6606 AccuPower Plus DualStar TM qpcr PreMix (with UDG), 50 μl/rxn, 8-tube strips, 96 rxn, ABI7500, optical film included K-6607 AccuPower Plus DualStar TM qpcr PreMix (with UDG), 50 μl/rxn, 8-tube strips, 96 rxn, Opticon, optical film included K-6608 AccuPower Plus DualStar TM qpcr Master Mix (2X) (with UDG), 2.5 ml, 100 rxn 86

AccuPower Dual-HotStart RT-qPCR PreMix & 2X Master Mix RT 와 qpcr 에 HotStart 기술을적용하여높은특이성 / 민감도로 One-step RT-qPCR Ease-of-use: 각각의 PCR tube에 RocketScript TM RTase, HotStart Taq DNA polymerase cdna 합성과 qpcr에필요한모든구성성분이포함되어있어실험자는 template RNA, primers & probe 및 D.W. 만첨가함으로써 Real-Time RT-PCR을수행할수있습니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system으로대량생산되어각 batch에대한철저한 QC를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. Description AccuPower Dual-HotStart TM RT-qPCR PreMix 는종래에비특이적으로일어나는역전사반응의문제점들을근본적으로개선한 HotStart 역전사기술이적용된제품입니다. 원하는 RNA 만선택적으로역전사하므로민감도가획기적으로개선되어극미량의 template RNA 에서도효과적으로합성할수있습니다. Template RNA 를대상으로하는각종바이러스검사및 gene expression 정량분석실험등에유용하게사용할수있는탁월한고감도 one-step RT-qPCR 제품입니다. Real-Time PCR 반응에필요한모든시약이혼합되어있어, template RNA 와 primer & probe 의첨가만으로손쉽게실험가능하며실험자 error 를최소화하여항상재현성있고정확한결과를얻을수있습니다. 각 tube 에 1 회사용분량이분주된 single use 제품으로써 master mix 제품의반복사용에따른 cross contamination 을방지할수있습니다. 또한, Exicycler TM 96 장비외에도타회사장비에최적화된전용 tube 및 plate 도제공가능합니다. Premix 뿐만아니라 2X Master Mix 제품으로도제공이가능합니다. Specifications Enzyme: RocketScript TM RTase, HotStart Taq DNA polymerase 5 to 3 exonuclease activity: Yes 3 to 5 exonuclease activity: No 3 - A Overhang: Yes RNase H activity: Yes Application Gene expression profiling Target RNA quantification Microbial detection Viral/bacterial pathogen load determination Storage Temperature -20 Accurate results! Excellent reproducibility! World-class technology! - HotStart RT + HotStart qpcr = Dual HotStart RT-qPCR Features and Benefits Sensitivity: 일반적으로검출이어려운극미량의 target template가고농도의 RNA에포함되어있는시료에서도검출이가능하며, 반응성이우수하여다양한종류의 PCR inhibitor를포함하고있는혈액, 토양시료의 template RNA에서도정확한 RT-qPCR 결과를얻을수있습니다. 10~10 10 copies까지 10 log의넓은 dynamic range 를갖습니다. Specificity: PyroHotStart RT 반응과 HotStart PCR을이용한세계최초 Dua-HotStart RT-qPCR 반응은정확히 target 유전자만을검출할수있도록최적화되었습니다 (Figure 1). Multiplexing: 여러종류의형광 dye (probe) 와호환성이뛰어나다양한 target gene을동시에검출할수있습니다. 다양한 Template RNA 검출가능 : RocketScript TM RTase가포함되어있어고온에서 RT 반응이가능하며 RNA 2차구조가강하게형성된 template RNA 등에서도유효한정량 RT-qPCR 결과를얻을수있습니다. Figure 1. Dual-HotStart TM Technology. www.bioneer.co.kr 87

AccuPower Dual-HotStart RT-qPCR PreMix & 2X Master Mix Experimental Data Figure 2. High sensitivity of AccuPower Dual-HotStart TM RT-qPCR PreMix. Experiment with HIV target. 10 fold serial dilution of template RNA (10 10 copies ~ 10 copies spiked in human total RNA). Figure 3. High specificity of AccuPower Dual-HotStart TM RT-qPCR PreMix. Experiment with HCV target. 10 fold serial dilution of template RNA (106 copies ~ 10 copies spiked in human total RNA). Conventional hotstart qpcr always generate non-specific amplification at low template concentration, which deteriorate the sensitivity of qpcr. AccuPower Dual-HotStart TM RT-qPCR PreMix accurately amplifies target RNA without non-specific amplification, even at low concentration of template Figure 4. Comparison of amplification quality between AccuPower Dual-Hotstart TM RT-qPCR PreMix and other competitor s master mix. Ordering Information Cat. no. Product Description K-6704 AccuPower Dual-HotStart TM RT-qPCR PreMix, 50 μl/rxn, 96-well plate, 96 rxn, Exicycler TM 96, optical film included K-6705 AccuPower Dual-HotStart TM RT-qPCR PreMix, 50 μl/rxn, 96-well plate, 96 rxn, ABI7500, optical film included K-6706 AccuPower Dual-HotStart TM RT-qPCR PreMix, 50 μl/rxn, 8-tube strips with cap, 96 rxn, Opticon, optical film included K-6707 AccuPower Dual-HotStart TM RT-qPCR Master Mix (2X), 100 rxn 88