01 Cloning AccuRapid Cloning Kit NEW 3 Gene Cloning Service 4 AccuPower Ligation PreMix 6 T4 DNA Ligase 7 Thermostable Thermus filiformis (Tfi) DNA Li
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1 Cloning DNA Amplification DNA Preparation Cloning (Cloning Kit & Gene Cloning Service) Phone: Cloning (Ligase) Phone: (ext.4->2) DNA Amplification Phone: (ext.4->2) DNA Preparation Phone: (ext.4->2)
2 01 Cloning AccuRapid Cloning Kit NEW 3 Gene Cloning Service 4 AccuPower Ligation PreMix 6 T4 DNA Ligase 7 Thermostable Thermus filiformis (Tfi) DNA Ligase 8
3 AccuRapid Cloning Kit NEW Features and Benefits 30분반응으로빠르고정확한 cloning Insert에제한효소를처리하지않고 vector에원하는방향으로삽입가능 Insert 양말단 complementary sequence 를인식하여연결되므로, PCR primer 디자인을통해여러조각 (1~3 조각 ) 삽입가능 Cloning 디자인을다양화하여원하는형태로 vector 조작 Application AccuRapid TM Cloning Kit 는 1~3 조각의 insert (PCR product) 를선형화된 vector 에정확하고신속하게 cloning 할수있는제품입니다. 본제품은 PCR 로증폭된 insert 말단과선형화된 vector 양말단의 18 ~ 21 bp complementary sequence 를인식하여연결하는방법으로, 기존처럼 insert (PCR product 또는 plasmid) 에제한효소를처리한후 ligation 하는방법이아니므로 vector 에 insert 를원하는방향으로삽입할수있습니다. Insert 증폭을위한 PCR primer 는선형화된 vector 양말단의 18 ~ 21 bp complementary sequence 를추가하여쉽게디자인할수있습니다. Multiple fragment cloning Gene synthesis Gene cloning Mutagenesis Vector modification Fusion protein Procedure Cat. no. Product Description K-7110 AccuRapid TM Cloning Kit, 10 rxns K-7120 AccuRapid TM Cloning Kit, 20 rxns K-7130 AccuRapid TM Cloning Kit, 50 rxns 3
4 Gene Cloning Service 바이오니아의오랜경험과풍부한분자생물학기술로번거롭고많은시일이소요되는 cloning 및 sequence 확인과정을한번에해결해드립니다. Features and Benefits 빠르고경제적인서비스 - 유전자원재료부터초고속올리고합성기까지일괄생산체계구축으로빠르고저렴한서비스. Gene 합성국내최저가 정확한품질 - Automatic DNA sequencer(abi 3730) 를이용한 100% sequence guarantee Application 단백질기능향상원하는 vector와의 cloning을통해단백질의발현효율을높일수있습니다. 항체유전자제작각종질병진단을위한항체제작시발현하고자하는숙주에코돈을최적화함으로써발현효율을높일수있으며, 또한고역가항체를얻기위해 diversity를극대화한여러형태의항체라이브러리를손쉽게얻을수있습니다. 유용물질생산생물체개발각종산업유용물질을생산하는균주들의유용물질생산에관련된유전자들을최적화하여생산효율을향상시킬수있습니다. Procedure 의뢰가능 Materials 서비스비용추가비용소요기간부가서비스 Gene Cloning Service Plasmid, PCR product, Cultured E. coli cell 200,000원 /cloning ( ~ 1 kb insert, ~ 7 kb vector) * Bioneer에서제공하는 vector에 cloning 가능 Cultured cell 이용시 50,000원추가 Insert 1 kb ~: 50,000원 /500 bp 7 ~ 10 kb: 100,000원추가 Vector 10 kb ~: Inquire Low copy: Inquire 1 ~ 8 kb (vector+insert) Average 5 ~ 10 working days 8 ~ 11 kb (vector+insert) Average 10 ~ 15 working days 11 kb ~ (vector+insert) Inquire Plasmid 증량서비스 (100,000원/100 μg) * Low copy plasmid 서비스제외 E. coli cell stock 제공서비스 (100,000원) 유전자합성과병행할경우 (100,000원할인 ) 유전자의구조, 특성등에의해가격및기간이상승될수있습니다. Commercial vector 를이용하여 cloning service 이용시 vector 구입비용이별도로청구됩니다. Template DNA 송부시, plasmid DNA 150~200 ng/μl, vol. 10 μl 이상, purified PCR product 50 ng/μl, vol. 10μl 이상을보내주셔야합니다. Sample 을잘못보내주실경우, 총비용의 50% 가추가청구되오니확인후보내주시기바랍니다. 진행중취소할경우금액의 50% 청구됩니다. 의뢰한 Cloning Service 를부득이하게 hold 할경우, 최대 1 개월이며 1 개월이후에는자동으로주문이취소되고금액의 50% 가청구됩니다. 4
5 Gene Cloning Service 주문방법 1. 바이오니아홈페이지 ( Gene Synthesis Service 페이지에서온라인주문하기배너를클릭하셔서주문페이지로들어갑니다. 2. 주문에서주문양식파일을다운받아작성하신후 3. 의뢰하신서비스를분석한후에서비스정보 ( 최종가격 + 기간 ) 를이메일로송부해드립니다. 4. 서비스의뢰를최종결정하시면이메일 (geneorder@bioneer. co.kr) 로통보해주십시오. 5. 서비스시작알림메일을보내드리는시점부터서비스가시작됩니다. Template DNA 송부방법박스에소속기관과의뢰자명을정확히기입하시고, 아래의당사연계배송회사를통하여 sample 을배송해주세요. 당사연계택배회사 : 현대택배 보내실곳 : 대전시대덕구문평서로 8-11 ( 주 ) 바이오니아 Synthetic Biology 팀 (Tel: , 8515) 운송비 : template 개수에상관없이당사부담 ( 단, 연계택배회사외에다른택배이용시운송비는고객께서부담하셔야합니다.) 주문취소규정주문후 5 일이내취소시금액의 50% 가청구됩니다. 주문후 5 일이후취소시금액의 80% 가청구됩니다 * Mutagenesis/Gene Cloning Service 는 material 수령시점부터서비스가시작됩니다. 상담안내 Tel: ( 고객지원센터 ) 또는 , 8515 (Synthetic Biology 팀 ) genesynthesis-support@bioneer.com 상담시간 : 평일오전 9:00 ~ 오후 6:00 ( 주말, 휴일휴무 ) 5
6 AccuPower Ligation PreMix 짧은시간에상온에서 Ligation Application Cloning into vectors Library construction TA cloning Linker ligation Recirclization of linear DNA Experimental Data AccuPower Ligation PreMix 는 T4 DNA Ligase, ATP 및반응버퍼등 ligation reaction 수행에필요한구성성분을혼합하여 0.2 ml tube 에동결건조시킨제품으로, 실온에서 5 분 (blunt-end DNA 는 10 분 ) 이내에 cohesive-end DNA 를 plasmid vector 에고효율로 ligation 할수있습니다. 또한바이오니아가독자개발한안정화물질이첨가되어있어실온에서도활성이 4 개월동안유지되며냉동 (-20 ) 보관을하실경우에는 3 년이상활성이지속적으로유지됩니다. Figure 1. 반응시간에따른 AccuPower Ligation PreMix의효율성실험. Lane 1 ~ 8: Lambda DNA / Hind III fragment (1 μg) Lane 9 ~ 16: Lambda DNA / EcoR V fragment (1 μg) Lane 2, 10: 60분동안 ligation Lane 3, 11: 50분동안 ligation Lane 4, 12: 40분동안 ligation Lane 5, 13: 30분동안 ligation Lane 6, 14: 20분동안 ligation Lane 7, 15: 10분동안 ligation Lane 8, 16: 5분동안 ligation Features and Benefits Fast Reaction Time: 실온에서 cohesive-end DNA 는 5 분, blunt-end DNA 는 10 분이내에 ligation 이가능합니다. Stability: 첨가된안정화제에의해실온에서는 4 개월, -20 에서는 3 년이상효소의활성이유지됩니다. Ease-of-use: T4 DNA Ligase, ATP, 최적화된반응버퍼를 1 회분량씩 0.2 ml tube 에포함하고있는제품으로 vector 와 insert DNA 만첨가해주면됩니다. Reproducibility: 재현성있는결과를위해바이오니아의전제품은엄격한 ISO 품질시스템하에서생산됩니다. Figure 2. 타사제품과의 ligation 효율비교. Lane 1, 9: Intact Lambda DNA (1 μg) Lane 2 ~ 8: Lambda DNA / Hind III fragment (1 μg) Lane 10 ~ 16: Lambda DNA / EcoR V fragment (1 μg) 3, 4, 11, 12: Bioneer AccuPower Ligation PreMix 5, 13: Company N T4 DNA ligase 6, 14: Company N Quick Ligation Kit 7, 15: Company P LigaFast Rapid DNA ligation system 8, 16: Company A Ready-To-Go T4 DNA ligase Cat. no. Product Description K-7103 AccuPower Ligation PreMix, 96 tubes, 0.2 ml 8-tube strips, 20 μl reaction 6
7 T4 DNA Ligase For Ligation of DNA, TA Cloning, and Other Recombinant DNA Applications Reagents Supplied 10 x Reaction Buffer: 500 mm Tris-HCI (ph 7.8), 100 mm MgCl 2, 50 mm DTT, 10 mm ATP, 25 μg/ml BSA Concentration 20,000 U (200 U/μl) Storage Conditions 50% glycerol containing 10 mm Tris-HCI (ph 7.5), 50 mm KCI, 1 mm EDTA, 10 mm 2-mercaptoethanol 본제품은유전자재조합실험에서 DNA 사이의결합에사용됩니다. T4 DNA ligase 는 duplex DNA 또는 RNA 에있는 5 - 인산기말단과 3 - 수산기말단부위사이에 phosphodiester bond 형성을촉진하여 duplex DNA, RNA, 또는 DNA/RNA hybrids 에있는단일가닥틈사이를 repair 할뿐만아니라 blunt-end 그리고 cohesive-end 끝말단부위로부터 DNA 를연결시킵니다. Features and Benefits Storage Temperature -20 Unit Definition 0.01 Weiss unit of enzyme is defined as the amount of enzyme required to give 90% ligation of Hind III fragments of lambda DNA in 30 min, at 16 in 20 μl of the assay mixture. Experimental Data High Speed: cohesive end DNA에서 25, 10분, blunt end DNA에서 25, 10분반응으로 DNA ligation이가능합니다. Flexibility: 대부분의 DNA ligation에최적화된제품입니다. Reproducibility: 재현성있는결과를위해바이오니아의전제품은엄격한 ISO 품질시스템하에서생산됩니다. Specifications Heat Inactivation: 70 for 10 min Application Blunt or cohesive-end ligation Repair of nicks in double-stranded nucleic acids Figure 1. T4 DNA Ligase를이용한 ligation. Lane 1: DNA fragment (digested with EcoR V) Lane 2, 3, 4, 5: T4 DNA Ligase 1 U, 16, 10, 20, 30 and 60 min Lane 6, 7, 8, 9: T4 DNA Ligase 1 U, 25, 10, 20, 30 and 60 min Lane 10, 11, 12, 13: T4 DNA Ligase 1 U, 37, 10, 20, 30 and 60 min Lane 14: Lambda DNA (digested with Hind III) Lane 15, 16, 17: T4 DNA Ligase 1 U, 16, 10, 20 and 30 min Lane 18, 19, 20: T4 DNA Ligase 1 U, 25, 10, 20 and 30 min Lane 21, 22, 23: T4 DNA Ligase 1 U, 37, 10, 20 and 30 min Cat. no. Product Description E-3061 T4 DNA Ligase, 20,000 U, 1 tube E-3062 T4 DNA Ligase, 100,000 U, 20,000 U x 5 tubes 7
8 Thermostable Thermus filiformis (Tfi) DNA Ligase Unit definition One unit of Tfi DNA Ligase is defined as the amount of enzyme required to give 50% ligation of the 12 base pair cohesive ends of 1 μg of PspE I digested lambda DNA in 10 min at 45. Tfi DNA Ligase는 DNA Ligase 의일종으로서절단된이중나선DNA 분자의인접한 5 -인산기말단과 3 -수산기말단사이를인산디에스테르결합 (phosphodiester bond) 으로연결해주는효소입니다. 특히다른 T4 DNA ligase, E. coli DNA ligase 등에비해더높은온도에서활성이안정하게유지되므로높은온도 (45~65 ) 에서반응할수있어반응시간이단축되고반응특이성이향상되었습니다. Source: Tfi DNA Ligase is isolated from E. coli cells containing the ligase gene cloned from Thermus filiformis. Application Ligase Chain Reaction (LCR) Oligonucleotide Ligation Assay (OLA) Mutagenesis by Incorporation of a phosphorylated oligo during PCR Amplification Simultaneous Mutagenesis of Multiple Sites Activity Assay Conditions The activity assay is carried out in a 20 μl reaction containing 1 μg of PspE I digested lambda DNA and 1 x Tfi DNA ligase reaction buffer. After incubation at 45 for 10 min., the reaction is terminated by addition of stop solution (40%(w/v) sucrose, 50 mm EDTA and 0.25% bromophenol blue). Then heat at 70 for 10 min and immediately load on a 0.8% agarose gel. Stability The half-life of the enzyme in 1 x reaction buffer is more than 1 hr at 95 and 55 hr at 65. Note: Tfi DNA Ligase should not be used as a substitute for other DNA ligase, i.e., T4 DNA Ligase. References Barany, F. (1991) Proc. Natl. Acad. Sci. USA, 88, Landegren, U. et al.(1988) Science 241, Michael, Scott F. (1994) Biotechniques 16:3, Gerard J. A. et al. (1993) Biotechniques 15:1, Reagents Supplied 10 x Reaction buffer (1 ml): 300 mm Tris-HCl (ph 8.3), 250 mm KCl, 50 mm MgCl 2, 5 mm NAD 1 x Dilution buffer (1 ml): 10 mm Tris-HCl (ph 7.6), 0.1 mm EDTA, 50 mm KCl, 1 mm DTT, 200 μg/ml acetylated BSA, 50% Glycerol Storage Condition 20 mm Tris-HCl (ph 7.6), 2 mm MgCl 2, 1 mm EDTA, 1 mm DTT, 0.5% Tween -20, 0.5% IGEPAL CA-630, 50% Glycerol, store at -20. Concentration 20 units/μl 8
9 Thermostable Thermus filiformis (Tfi) DNA Ligase Experimental Data Heat Stability test at 95 C and 65 C Fragment 4. Fragment Ligation Product (1+4) Figure 1. Ligation test at various temperatures (45 ~65 ) Incubate the reaction containing ligase 1 unit and 1 μg DNA[lambda PspE I] at each temperature for 10 min. Lane 1: λdna/pspe I (control) Lane 2: Incubate at 45, 10 min Lane 3: Incubate at 50, 10 min Lane 4: Incubate at 55, 10 min Lane 5: Incubate at 60, 10 min Lane 6: Incubate at 65, 10 min Figure 2. Heat Stability test at 95. Incubate the enzyme at 95 each time. And then add 1 unit ligase to a 20 μl reaction containing 1 μl DNA[lambda PspE I] and incubate the mixture at 45 for 10 min. Lane 1: λdna/pspe I (control) Lane 2: Incubate at 95, 10 min Lane 3: Incubate at 95, 20 min Lane 4: Incubate at 95, 30 min Lane 5: Incubate at 95, 40 min Lane 6: Incubate at 95, 50 min Lane 7: Incubate at 95, 60 min Lane 8: Incubate at 95, 70 min Lane 9: Incubate at 95, 80 min Lane 10: Incubate at 95, 90 min Cat. no. Product Description E-3111 Tfi DNA Ligase, 2,000 U, 10 x reaction buffer, 1 ml, 1 x dilution buffer, 1 ml E-3112 Tfi DNA Ligase, 10,000 U, 10 x reaction buffer, 1 ml, 1 x dilution buffer, 1 ml 9
10 02 DNA Amplification AccuPower Pfu PCR PreMix 11 AccuPower ProFi Taq PCR PreMix 13 AccuPower HotStart Pfu PCR PreMix 15 Pfu DNA Polymerase 17 ProFi Taq DNA polymerase 19
11 AccuPower Pfu PCR PreMix Gene Cloning 등 High Fidelity 를요구하는실험에최적화된 Kit 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. 11
12 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) 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 12
13 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 AccuPower ProFi Taq PCR PreMix 는바이오니아에서독자적으로개발한 ProFi Taq DNA polymerase 와 PCR 반응혼합물이진공건조된제품으로, template DNA, primer 그리고 D.W. 의첨가만으로정확하고깨끗한 PCR 산물을얻을수있는제품입니다. 본제품에사용되는 ProFi Taq DNA polymerase 는높은정확성과뛰어난증폭효율을나타내며, Human Genomic DNA 의경우최대 21 kb (Lambda DNA 의경우최대 30 kb) 의 DNA fragment 증폭이가능합니다. AccuPower ProFi Taq PCR PreMix 는 high efficiency 및 longrange PCR 에매우적합한제품이며, 그외에도다양한 PCR 에사용하실수있습니다. 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. 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 Supplier I Supplier S Supplier T 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) Stability: PCR 반응혼합물에안정화제를첨가하여건조시켜, 장기간보관하더라도활성을안정적으로유지합니다. Reproducibility: ISO 9001 품질시스템하에서 one-batch system 으로대량생산되어각 batch 에대한철저한 QC 를거친후균일한품질의제품으로공급되기때문에사용자가대량의시료를반복적으로처리할때각 tube 마다발생하는편차문제를해결하여재현성있는실험결과를제공합니다. 13
14 AccuPower ProFi Taq PCR PreMix M Supplier I Supplier S Supplier T M M1M Supplier I Supplier S Supplier T 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) 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 14
15 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 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. 15
16 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) 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 16
17 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 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: 300 mm Tris-HCl, 200 mm KCl, 100mM (NH 4) 2SO 4, 15 mm MgSO 4, Acetylated BSA, ph 9.0 Dilution buffer: 50 mm Tris-HCl, 0.1 mm EDTA, 1 mm DTT, Stablizers, 50% Glycerol, ph 8.0 dntps mixture: 10 mm, each dntp 2.5 mm (optional) Concentration 250 U (2.5 U/μl) 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. 17
18 Pfu DNA Polymerase Experimental Data M M 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) Cat. no. Product Description E-2015 Pfu DNA Polymerase, 250 U, 10 x reaction buffer E Pfu DNA Polymerase, 250 U, 10 mm dntps, 10 x reaction buffer E-2016 Pfu DNA Polymerase, 1,000 U, 10 x reaction buffer 18
19 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 ProFi Taq DNA Polymerase 는합성생물학기술을이용하여개발한 recombinant Taq DNA polymerase 로써뛰어난증폭효율과정확성을동시에충족시키는 DNA polymerase 입니다. ProFi Taq DNA polymerase 는 Taq DNA polymerase 를개선한효소로써기존의 Taq DNA polymerase 와비교하여뛰어난증폭성능, 높은 efficiency 를가지는제품입니다. 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. Reagents Supplied 10 x Reaction Buffer: 400 mm Tris-HCl, 600 mm KCl, 15 mm MgCl 2, Acetylated BSA, ph 9.0 Dilution buffer: 20 mm Tris-HCl, 0.5 mm EDTA, 1 mm DTT, 100 mm KCl, Stablizers, 50% Glycerol, ph 8.0 dntps mixture: 10 mm, each dntp 2.5 mm Concentration 250 U (5 U/μl) Storage Temperature -20 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. 19
20 ProFi Taq DNA Polymerase Experimental data Bioneer M Supplier T Supplier S Supplier I 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) Bioneer M1 M Supplier I Supplier S Supplier T M2 M1 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) M1M Supplier I Supplier S Supplier T M2M1 Figure 3. 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, 30 cycles of 95 for 20 sec, 65 for 20 sec, and 68 for 4 min. PCR reactions using other suppliers DNA polymerase were performed according to each supplier s protocol. Lambda DNA was used as a template for PCR amplification. Lane 1: 15 kb fragment (human tumor protein p53 gene) Lane 2: 20 kb fragment (human tumor protein p53 gene) Lane 3: 25 kb fragment (human DNA cross-link repair 1A gene) Lane 4: 30 kb fragment (human hemoglobin epsilon 1 gene) Lane M1: Lambda/Hind III marker (Bioneer, Cat. no. D-1050) Lane M2: 1 kb DNA Ladder (Bioneer, Cat. no. D-1040) Cat. no. Product Description E-2201 ProFi Taq DNA Polymerase 250 U, 10 mm dntps, 10 X reaction buffer with MgCl 2 E-2202 ProFi Taq DNA Polymerase 250 U, 10 mm dntps, 10 X reaction buffer without MgCl 2, 20 mm MgCl 2 E-2203 ProFi Taq DNA Polymerase 250 U, 10 X reaction buffer with MgCl 2 E-2204 ProFi Taq DNA Polymerase 250 U, 10 X reaction buffer without MgCl 2, 20 mm MgCl 2 E-2205 ProFi Taq DNA Polymerase 1000 U, 10 mm dntps, 10 X reaction buffer with MgCl 2 E-2206 ProFi Taq DNA Polymerase 1000 U, 10 mm dntps, 10 X reaction buffer without MgCl 2, 20 mm MgCl 2 E-2207 ProFi Taq DNA Polymerase 1000 U, 10 X reaction buffer with MgCl 2 E-2208 ProFi Taq DNA Polymerase 1000 U, 10 X reaction buffer without MgCl 2, 20 mm MgCl 2 20
21 03 DNA Preparation Magnetic Bead Type Kit MagListo 5M Plasmid Extraction Kit 21 MagListo 5M PCR Purification Kit 23 MagListo 5M Gel Extraction Kit 24 Spin Column Type Kit AccuPrep Nano-Plus Plasmid Mini Extraction Kit 25 AccuPrep Plasmid Mini Extraction Kit 27 AccuPrep PCR Purification Kit 29 AccuPrep Gel Purification Kit 32
22 MagListo 5M Plasmid Extraction Kit Plasmid Mini, Midi and Maxi Prep 을한번에! 빠르고간편한신개념 Plasmid DNA 추출키트 Experimental Data MagListo TM 5M Plasmid Extraction Kit 은 Magnetic Nanobead 와 MagListo TM Magnetic Separation Rack 을이용하여 5 분 (mini), 10 분 (midi), 15 분 (maxi) 내에배양된박테리아세포로부터 plasmid DNA 를추출할수있는제품입니다. 자성분리방식으로세포분쇄물과 plasmid DNA 를분리, 농축및정제하는과정을거치기때문에원심분리기를사용하는방법에비해빠르게 plasmid DNA 를분리할수있습니다. 본제품은한 kit 를사용하여 mini, midi, maxi scale 로 plasmid DNA 를추출할수있어별도의 kit 를구매할필요가없으며, vacuum system 이나 air pressure system 을구비할필요가없어실험비용을절감할수있습니다. Figure 1. Electrophoresis data of 500 ng of several size plasmids purified with MagListo TM 5M Plasmid Extraction Kit. M1: Bioneer 100 bp ladder 1: 3.5 kb plasmid DNA 2: 5.4 kb plasmid DNA 3: 8 kb plasmid DNA M2: Bioneer 1 kb ladder Features and Benefits Magnetic Nanobead를이용하여 mini-5분, midi-10분, maxi- 15분에 plasmid 추출 Mini, midi, maxi prep kit을따로구매할필요없이하나의 kit 로해결 Extraction kit와별도의 MagListo TM rack 외에고가의추가장비불필요 Application Gene cloning, PCR, Real-Time PCR, sequencing, transformation, transfection, in vitro transcription/translation Specifications Figure 2. Comparison of plasmid DNAs purified using MagListo TM and competitors products (magnetic bead type). M1: Bioneer 100 bp ladder 1-2: Plasmid DNA purified with Bioneer MagListo TM (3.5 kb / 5 kb) 3-4: Plasmid DNA purified with competitor P kit (3.5 kb / 5 kb) 5-6: Plasmid DNA purified with competitor I kit (3.5 kb / 5 kb) M2: Bioneer 1 kb ladder 1(B) 2(B) 3(B) 4(B) 5(B) 6(B) Concentration(ng/ul) A260/ Mini scale Midi scale Maxi scale Starting culture volume 1~5 ml 20~50 ml 100~200 ml Elution volume 100 μl 500 μl 1 ml Expected DNA yield ~ 20 μg ~ 400 μg ~ 1 mg Preparation time ~ 5 min ~ 10 min ~ 15 min 22
23 MagListo 5M Plasmid Extraction Kit Figure 3. Comparison of plasmid transfection efficiency in 293T cells between MagListo TM and a competitor s product (spin column type). 293T cell 1x10 6 을이용한 transfection efficiency 확인결과, Bioneer MagListo TM 와경쟁사제품에서추출한 plasmid 모두 95% 이상의효율을확인할수있습니다. Figure 4. Comparison cell viability after transfected with prep samples by MagListo TM and a competitor s product (spin column type). 293T 세포주에 transfection 후 WST assay를시행한결과, plasmid DNA 400 ng/well (6 well plate) 농도까지 cell viability에영향을미치지않음을확인할수있습니다. Procedure Cat. no. Product Description K-3600 MagListo TM 5M Plasmid Extraction Kit, 500 rxn in mini K-3601 MagListo TM 5M Plasmid Extraction Kit, 100 rxn in mini 23
24 MagListo 5M PCR Purification Kit PCR Product 를비롯한다양한효소반응물의 Fragment DNA 정제를빠르고간편하게! Specifications Size range 100 bp ~ 10 kb Typical recovery 90 ~ 100% Expected purity A260/280 > 1.8, A260/230 > 1.6 Purification time ~ 4 min Application Subcloning, Sequencing, Labeling, DNA concentration and other molecular biological applications Experimental Data MagListo TM 5M PCR Purification Kit 는 Magnetic Nanobead 와 MagListo TM Magnetic Separation Rack 을이용하여 PCR product 를비롯한다양한효소반응물 ( 제한효소반응, A-tailing 반응, labeling 반응등 ) 에서 fragment DNA 를빠르게추출할수있는제품입니다. 자성분리방식을이용하여원심분리기를사용하는방식보다각종불순물 (dimer, salts, dntps, enzymes, mineral oil, ethidium bromide, dye, detergent 등 ) 들을손쉽고빠르게제거하여고순도의 fragment DNA 를정제할수있습니다. A B Features and Benefits MagListo TM Magnetic Separation Rack를이용하여 4분만에 DNA 정제 원심분리기등의고가장비불필요 Figure 1. Comparison of fragment DNA purified with MagListo TM PCR Purification Kit and control. A. C: Control 100 bp DNA Ladder (D-1030) 1-2: 100 bp DNA Ladder purified with MagListo TM 5M PCR Purification Kit B. C: Control 1 kb DNA Ladder (D-1040) 1-2: 1 kb DNA Ladder purified with MagListo TM 5M PCR Purification Kit Procedure Cat. no. Product Description K-3609 MagListo TM 5M PCR Purification Kit, 100 rxn in mini 24
25 MagListo 5M Gel Extraction Kit Agarose Gel 로부터 Fragment DNA 정제를빠르고간편하게! Specifications Size range 100 bp ~ 10 kb Typical recovery 90 ~ 100% Expected purity A260/280 > 1.8 Purification time ~ 15 min Application Subcloning, Sequencing, Labeling, DNA concentration and other molecular biological applications Experimental Data MagListo TM 5M Gel Extraction Kit 는 Magnetic Nanobead 와 MagListo TM Magnetic Separation Rack 을이용하여 TAE, TBE agarose gel 에서원하는 fragment DNA 를추출할수있는제품입니다. Gel Solubilization Buffer 에는 ph indicator 가함유되어있어최적의 binding 조건을잡을수있으며, 자성분리방식을이용하여원심분리기를사용하는방법에비해빠르게고순도의 fragment DNA 를정제할수있습니다. Features and Benefits Magnetic Nanobead를이용하여 15분만에 agarose gel로부터 DNA 정제 원심분리기등의고가의장비불필요 Figure 1. Comparison of fragment DNA from TBE agarose gel purified with MagListo TM 5M Gel Extraction Kit and competitor s kit (spin column type). M: Bioneer 1 kb ladder 1-2: 500 bp DNA purified with Bioneer MagListo TM 5M Gel Extraction Kit 3-4: 500 bp DNA purified with competitor Q kit 5: Control 500 bp DNA 6-7: 3 kb DNA purified with Bioneer MagListo TM 5M Gel Extraction Kit 8-9: 3 kb DNA purified with competitor Q kit 10: Control 3 kb DNA 11-12: 5 kb DNA purified with Bioneer MagListo TM 5M Gel Extraction Kit 13-14: 5 kb DNA purified with competitor Q kit 15: Control 5 kb DNA Procedure Cat. no. Product Description K-3607 MagListo TM 5M Gel Extraction Kit, 100 rxn in mini 25
26 AccuPrep Nano-Plus Plasmid Mini Extraction Kit Specifications Starting culture volume Column binding capacity Elution volume Expected DNA yield Preparation time 1 ml ~ 10 ml > 20 μg μl ~ 20 μg ~ 10 min Application Subcloning, Sequencing, Transformation, Transfection, In vitro transcription/translation AccuPrep Nano-Plus Plasmid Mini Extraction Kit 는단 10 분내에고순도, 고농도의 plasmid DNA 를추출할수있는제품으로써 Birnboim 의 alkaline lysis method 와바이오니아의독창적인세포잔해입자및단백질제거특허기술이결합된신개념제품입니다. 본제품에포함된나노입자는용액내 insoluble protein aggregate 및 chromosomal DNA, cell debris 등과함께 complex 를형성하여원심분리시매우빠른시간내에 precipitate 를형성할수있게해줍니다. 또한, 용액내포함된나노입자에의해 insoluble complex ( 나노입자 + protein aggregate + chromosomal DNA + cell debris) 가매우단단하게 tube 벽면에고정됨으로써기존실험방법에비해고순도의 cleared lysate 를얻을수있습니다. 본제품은 cleared lysate 를얻기위해수행하던원심분리시간을획기적으로단축시켜 (10 분 1 분 ) 단 10 분이내에전체실험진행이가능합니다. 또한, DNA binding capacity 가극대화된 silica based DNA binding column 을적용하여높은수율을보장합니다. Experimental Data Figure 1. Reduced total preparation time. B: AccuPrep Nano-Plus Plasmid Mini Extraction Kit Q, P: Competitors s plasmid extraction kit Features and Benefits 바이오니아의독창적세포잔해입자및단백질제거특허기술이도입된신개념 plasmid DNA 추출키트 단 10분이내에 E. coli cell로부터고순도, 고수율의 plasmid DNA 추출 High-copy 및 low-copy plasmid DNA에도모두사용가능 enda+ strain을위한 Endonuclease A denaturation buffer 제공 (Denaturation Buffer, PD) 높은 binding capacity를갖는 silica based DNA binding column 사용 Figure 2. Restriction enzyme digestion test. AccuPrep Nano-Plus Plasmid Mini Extraction Kit를이용하여추출한 plasmid DNA를여러종류의제한효소로 digestion한후 agarose gel에서확인하였습니다. Left- pbluescript SK(+) Right- pbi121 Lane M: Size marker(cat. no. D-1040, Bioneer) 26
27 AccuPrep Nano-Plus Plasmid Mini Extraction Kit Procedure Cat. no. Product Description K-3111 AccuPrep Nano-Plus Plasmid Mini Extraction Kit, 200 rxn K-3112 AccuPrep Nano-Plus Plasmid Mini Extraction Kit, 50 rxn KB-0101 RNase A powder, lyophilized (6 mg/tube) KA DNA Binding Column Tubes (50 ea X 4 box) 27
28 AccuPrep Plasmid Mini Extraction Kit Specifications Starting culture volume Column binding capacity Elution volume Expected DNA yield Preparation time 1 ml ~ 10 ml > 20 μg 50 ~ 100 μl ~ 20 μg ~ 20 min Application Subcloning, Sequencing, Transformation, Transfection, In vitro transcription/translation AccuPrep Plasmid Mini Extraction Kit 는고순도, 고농도의 plasmid DNA 를추출할수있도록개발된제품으로써 Birnboim 의 alkaline lysis method 에기초한제품입니다. 본제품은높은 DNA binding capacity 를가지고있는 silica based filter 가사용되어큰 size 의 plasmid DNA 나많은양의 plasmid DNA 추출에매우효과적으로사용할수있습니다. Experimental Data Features and Benefits 높은 binding capacity를갖는 silica based DNA binding column 사용 enda+ strain을위한 Endonuclease A denaturation buffer 제공 (Denaturation Buffer, PD) DNA binding column 및 collection tube가들어있는 50 well tube rack은추출된 DNA 보관용 tube rack으로활용가능 Figure 1. Restriction enzyme (Xba I) digestion test. 200 ng of extracted plasmid DNA (pbluescript SK(+)) was digested with Xba I. Lane M: Molecular weight marker (λdna/hind III+EcoR I, Cat. no. D-1070, Bioneer) Lane 1: AccuPrep Plasmid Mini Extraction Kit Lane 2, 3: competitor s plasmid extraction kit Figure 2. Restriction enzyme digestion test. 200 ng of extracted plasmid DNA (pbluescript SK(+)) was digested with various restriction enzymes. Lane M: Molecular weight marker (λdna/hind III+EcoR I, Cat. no. D-1070, Bioneer) K: Kpn I, X: Xho I, SI: Sal I, H: Hind III, E: EcoR I, P: Pst I, Sm: Sma I, B: BamH I, S: Spe I, Xb: Xba I, N: Not I, Sc: Sac I 28
29 AccuPrep Plasmid Mini Extraction Kit Procedure Cat. no. Product Description K-3030 AccuPrep Plasmid Mini Extraction Kit, 200 rxn K AccuPrep Plasmid Mini Extraction Kit, 50 rxn KB-0101 RNase A powder, lyophilized (6 mg/tube) KA DNA Binding Column Tubes (50 ea X 4 box) 29
30 AccuPrep PCR Purification Kit Application Subcloning, Sequencing, Labeling, DNA concentration, etc. Experimental Data AccuPrep PCR Purification Kit 는 PCR 반응물을비롯한다양한효소반응물 ( 제한효소반응, A-tailing 반응, labeling 반응등 ) 로부터 5 분이내에각종불순물 (dimer, salts, dntps, enzymes, mineral oil, ethidium bromide, dye, detergent 등 ) 을완벽히제거하여고순도의 fragment DNA 를정제할수있는제품입니다. 본제품은 silica based DNA binding column 이사용된 spin column 방식의제품으로 binding buffer 와 washing buffer 및 elution buffer 로구성되어있습니다. 또한 elution volume 을 30 μl 까지줄일수있으므로 PCR product 농축에도효과적입니다. 정제된 fragment DNA 는 subcloning, sequencing, Southern blotting, Northern blotting 등에사용될수있습니다. Features and Benefits 100 bp부터 10 kb의 double strand DNA 및 single strand DNA 정제 평균 recovery는 70-90% 이상 전체실험소요시간이 5분미만으로매우빠르게실험가능 유기용매를사용하지않아매우안전하며, ethanol precipitation 과정이없어사용이편리함 recovery (%) B Q 200 bp 400 bp 1 kb 3 kb 6 kb Size of DNA fragment Figure 1. DNA size에따른 recovery 분석. 200 bp부터 6 kb까지의 fragment DNA를이용하여 recovery를분석한결과입니다. 평균 80% 이상의 recovery를보입니다. B: AccuPrep PCR Purification Kit Q: Competitor Q 1.0 kb M B B B Figure 2. Recovery analysis of the purified PCR product. 정제된 PCR product의정제효율을확인한결과입니다. Lane M: Molecular weight marker, 100 bp Plus ladder (D-1035, Bioneer) Lane 100: 100%, Lane 90: 90%, Lane 80: 80% Lane 70: 70%, Lane 60: 60%, Lane 50: 50% Lane B: purified PCR product 30
31 AccuPrep PCR Purification Kit Procedure Cat. no. Product Description K-3034 AccuPrep PCR Purification Kit, 200 rxn K AccuPrep PCR Purification Kit, 50 rxn KA DNA Binding Column Tubes (50 ea X 4 box) 31
32 AccuPrep Gel Purification Kit Specifications Subcloning, Sequencing, Labeling, DNA concentration, etc. Experimental Data AccuPrep Gel Purification Kit 는 Low melting agarose gel 뿐만아니라 TAE, TBE agarose gel 에서도원하는 DNA fragment 를 15 분이내에분리정제할수있도록고안된제품입니다. 또한, binding buffer 에 ph indicator 가함유되어있어최적의 binding 조건을잡을수있습니다. 본제품은 silica based DNA binding column 이사용된 spin column 방식의제품으로, 최적화된 binding buffer 와 washing buffer 및 elution buffer 가포함되어있어편리하게사용할수있습니다. 또한 elution volume 을 30 μl 까지줄일수있으므로 DNA 농축에도효과적입니다. Features and Benefits Low melting agarosge gel및 TAE/TBE agarose gel로부터 fragment DNA 추출 70 bp에서 10 kb 크기의 double strand DNA 및 single strand DNA 정제 전체실험소요시간이 15분미만으로매우빠르게실험가능 유기용매를사용하지않아안전하며, ethanol precipitation 과정이없어사용이편리함 recovery (%) B Q 70 bp 200bp 400bp 1 kb 3 kb 6 kb Size of DNA fragment Figure 1. DNA size에따른 recovery 70 bp부터 6 kb까지의 fragment DNA를이용하여 recovery를분석한결과입니다. B: AccuPrep Gel Purification Kit Q: Competitor Q 1.0 kb M B B B Figure 2. Recovery analysis of the purified fragment DNA. Agarose gel로부터 10.0 kb의 plasmid DNA를정제한후정제효율을확인한결과입니다. Lane M: Molecular weight marker, 100 bp Plus ladder (D-1035, Bioneer), Lane 100: 100%, Lane 90: 90%, Lane 80: 80% Lane 70: 70%, Lane 60: 60%, Lane 50: 50% Lane B: AccuPrep Gel Purification Kit 32
33 AccuPrep Gel Purification Kit Procedure Cat. no. Product Description K-3035 AccuPrep Gel Purification Kit, 200 rxn K AccuPrep Gel Purification Kit, 50 rxn KA DNA Binding Column Tubes (50 ea X 4 box) 33
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DNA technology (DNA 조작기술 ) 유전자 cloning Cloning & Plasmid Joshua Ledergerg & Edward Tatum, 1946 두종류의다른유전자를가진대장균사이에유전자의재조합이일어나새로운대장균이만들어지는것을발견했다. 이원리를바탕으로 1970 년대에 DNA 재조합기술이발전하게되었다. 원핵생물에서 DNA 의이동방법 플라스미드를이용한
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