Ver.0 (KR) 20190410 Next Generation Sequencing Service Bioneer Corporation is Korea s leading biotech company. Bioneer is the first Korean biotechnology company when it was established in 1992.
생명과학국내 1 위기업 BIONEER s Next Generation Sequencing Service NGS(Next Generation Sequencing) 은 Sanger Sequencing 과달리대량의병렬데이터생산 (Massive parallel sequencing) 으로유전체의염기서열을고속으로분석 (Highthroughput sequencing) 하는기술로, 하나의유전체를수많은조각으로분해하여각조각을동시에읽은후전산기술을이용하여조합함으로써방대한유전체정보를빠르게해독하는방법입니다. 바이오니아의 NGS Service 는연구자들에게연구목적에맞는다양한 NGS application 을제공합니다. 유전체 (Whole Genome), 엑솜 (Exome), 전사체 (Transcriptome), 후성유전체 (Epigenome) 에대한유전체해독및분석을제공합니다. 인간 (Human) 뿐만아니라다른어류및동 / 식물, high GC contents 비율의박테리아등다양한생명체에대한유전체해독, 분석을포함하여 de novo 나 reference 기반유전체해독모두가능합니다. 1
BT IT BIONEER CORPORATION NT 바이오니아는 BT, IT, NT 기술을융합한원천특허들을보유하고있는유전자전문기업으로, 분자진단및핵산치료제연구개발과함께여러전문가들과의지속적인협업을통해생명공학연구자들에게 Total solution 을제공을목표로합니다. 2
BIONEER s Next Generation Sequencing Service Application 게놈 - 엑솜해독 / 분석 전장유전체해독 De novo 해독엑솜해독타겟영역해독 미생물게놈 해독 / 분석 박테리아게놈해독메타지노믹스해독균류 ( 곰팡이, 진균 ) 게놈해독메타 16S rdna 해독 3
다양한 NGS 응용서비스제공연구프로젝트맞춤컨설팅신뢰도높은 data 제공 Basic/Customized Bioinformatics 분석연계서비스 : Sanger Sequencing, mrna Profiling 후성게놈해독 / 분석 Methyl 해독 Bisulfite 해독 MeDIP 해독 ChIP 해독 전사체해독 / 분석 전사체해독 Small RNA 해독 4
BIONEER s Next Generation Sequencing Service Workflow 접수된샘플들은각조건에맞추어 QC 가이루어지며, QC 후 Sequencing 진행여부를결정합니다. 약 2~3 일소요되며, DNA/RNA 추출서비스를추가요청하신경우기간이연장될수있습니다. 샘플 QC 주문서접수 샘플접수 1. 홈페이지 (www.bioneer.co.kr) 에서주문서를다운로드받습니다. 2. 고객및샘플정보, 프로젝트에대한세부내역을기재합니다. 3. NGS_support@bioneer.co.kr 로보냅니다. 상담및견적발행 영업사원이직접방문하거나택배를이용하여샘플을배송합니다. 1. 주문서확인후분석방법을논의합니다. ( 샘플특성, 플랫폼, 데이터생산량, 분석방법등 ) 2. 논의사항을바탕으로견적서를발행해드립니다. 5
Library Construction 각분석항목에맞추어 Library 를제작하는과정입니다. Sequencing 적절한플랫폼에 Sequencing 을진행하며, Library Construction 부터약 4~6 주소요됩니다. Data Analysis 요청하신분석항목에따라분석이진행되며기본분석기준약 1 주일소요됩니다. 최종 Data 및 Report 제공 주문서작성시기재된 E-mail 로 Raw data 및 Report 가발송됩니다. 6
Whole Genome Sequencing Whole Genome Sequencing (WGS) 은 model organism 과 human 의유전체전체를한번에읽어내어유전정보를분석하는방법입니다. WGS 를통해얻어진 Data 를활용하여 single nucleotide polymorphisms(snps), insertions/deletions(indels), copy number variants(cnvs) 그리고 Structural Variation 분석등변이분석이가능합니다. 생물정보학도구를이용해분석된 data 는 report 와함께 Variant Call Format(VCF), Annotation Result Excel Format(xls) 이제공됩니다. Resequencing 이미알려진 reference genome 을이용해유전적변이를확인하고비교하는데유용합니다. Guide Line Objective Individual Genome/ General Disease Obtaining variants of each sample for downstream analysis Population Genomics Population and phylogenetic analysis disease and phenotype relationship analysis Cancer / Rare Disease Detecting cancer specific and/or rare variants Sufficient depth 30X 10X 100X/200X Sample Requirements Minimum Quantity 1 µg, Minimum Concentration = 50 ng/µl, OD 260/280=1.6~2.0 Deliverables Raw data(fastq), Statistics of raw data, Mapping to reference genome, SNPs and InDels calling, Variant annotation, Functional annotations * 샘플 type, 종, data analyses 에따라변경될수있습니다. De novo sequencing Reference 가없는새롭게발견된생물종의유전체정보를분석하는방법으로세밀한변종특성, 가변적인 genome 의비교연구를할수있습니다. De novo sequencing 은고난도의기술이요구되며 draft map/fine map 두가지기술이수행됩니다. 성공적인 de novo genome 을완성하기위해다양한 size 의 library(200 bp, 500 bp, 2 kb, 5 kb, 10 kb, 20 kb) 를만들어분석에이용합니다. Guide Line General sequencing depth Gene coverage Draft Map 60X 95% Fine Map 80X 98% Genome Coverage 90% (except heterochromatin region) 95% (except heterochromatin region) Accuracy Sample Requirements Minimum Quantity 3 µg, Minimum Concentration = 50 ng/µl, OD 260/280=1.6~2.0 Contig Sequence N50 Scaffold Sequence N50 99.999% >50 kb >20 kb 99.999% >20 kb >300 kb Deliverables Raw data(fastq), statistics of raw data, de novo assembly, taxonomy profiling, k-mer analysis, Gene prediction, Gene annotation * 샘플 type, 종, data analyses 에따라변경될수있습니다. 7
Whole Exome Sequencing Whole Exome Sequencing (WES) 은전체유전체의 1% (30 Mb), exon 이라는유전정보를담고있는 protein coding 영역만선택적으로분석하는방법입니다. 이영역을 specific kit 를이용하여선택적으로 capture 후 sequencing 을수행하기때문에 sequencing depth 도증가될수있고, whole genome sequencing 보다경제적입니다. 주로임상연구나진단, 예측적으로관련된유전자나변형을식별하기위해법의학에서도사용됩니다. Guide Line Whole Exome Sequencing Target Region Sequencing Objective Accurate variants detection within exon region Detecting cancer specific and/or rare variants Sufficient depth (human exome) Capture region 100X Agilent SureSelect Exome Capture Kit/ Customized Capture Kit Sample Requirements Minimum Quantity 1 µg, Minimum Concentration = 50 ng/µl, OD 260/280=1.6~2.0 500X Deliverables Raw data(fastq/vcf), Summary of data production, Mapping statistics, Statistics of sequencing reads, SNPs and InDels calling, Variant annotation, SNVs concordance, Tumor-Normal paired analysis, Trio(Family-based) analysis * 샘플 type, 종, data analyses 에따라변경될수있습니다. Exome/Targeted DNA Sequencing Process Construct shotgun library Hybridization Genomic DNA Fragments Pulldown Wash AGGTCGTTACGTACGCTAC GACCTACATAGCAGTTCAC GCATCACAAAGCTAGGTGT GACCTACATAGCAGTTCAC Mapping, alignment, variant calling DNA sequencing Captured DNA 8
Transcriptome Sequencing RNA sequencing(transcriptome) 을통해분석된정보는특정시점의활성된유전자정보를정량화및구조 / 기능적인분석을할수있습니다. RNA 자체기본적인연구는물론, 의학연구, 약물유전학, 맞춤형의학연구에활용될수있습니다 (gene expression profiles, post-translational modifications, SNPs or mutations over time). Total RNA, small RNA(miRNA, trna), de novo sequencing 모두가능합니다. 연계서비스 RNA Profiling Service: qpcr array service 로유전자탐색및검증까지 Guide Line Objective Reference Based RNA expression analysis gene structure prediction-alternative sliced transcriptome De novo Novel gene detection comparative expression analysis gene structure prediction Sufficient data quantity Raw data 4 Gb Raw data 6 Gb Sample Requirements Deliverables Minimum Quantity 1 µg, Minimum Concentration = 65 ng/µl, OD 260/280=1.6~2.0, RIN 7, 28S:18S>1.0 Mapping statistics, SNPs and InDels calling, Gene expression profiles, Differentially Expressed Gene(DEGs), Variant annotation, Fusion gene, Identification of alternative spliced transcripts, Prediction of novel transcripts * 샘플 type, 종, data analyses 에따라변경될수있습니다. De novo Assembly statistics, Gene annotations, Gene expression profiles, Differentially Expressed Gene(DEGs), Gene ontology analysis, Comparative analysis 9
Epigenome Sequencing 생물이가지는모든후성학적인유전자발현조절기전은 DNA metylation, 히스톤변형을통해조절되며, 유전되기도하고음식, 습관, 환경에의해영향을받는다고알려져있습니다. 후성학적인유전자변형은암을포함하여다양한질병들의발병및진행에관여한다고연구되고있으며, 이를분석한 data 를이용하여진단및수술, 처치에대한반응상태와질병진행을예측하는데유용하게쓰일수있습니다. High resolution 의장점을가지는 CHIP-Seq, MeDIP-Seq 및 Whole genome 의전체 Bisulfite-Seq 을이용하여분석하실수있습니다. Chromosome DNA methylation ME Methyl marks added to certain DNA bases repress gene activity. Histone modification Chemical Tag Histone ME Histone tail ME A combination of different molecules can attach to the tails of proteins called histones. These alter the activity of the DNA wrapped around them. DNA Metagemone Sequencing 단일생물종을넘어서서특정환경에서수집한샘플에함유된유전체를분석함으로써각환경의미생물군집을패턴차이및상호작용에대해확인하는방법입니다. 종식별마커로사용되는 16S rrna 나 18S rrna 등을통해최근주목받고있는장내미생물의분포및관련된임상연구, 생명공학등에널리사용될수있습니다. Normal Obese Innermost ring: phylum level Middle ring: family level Outermost ring: genus level Hu HJ et al., PLOS ONE, 2015 Bacteroidetes Bacteroidaceae Bacteroides Proteobacteria Rikenellaceae Alistipes Firmicutes Porphyromonadaceae Parabacteroides Others Prevotellaceae Prevotella Sutterellaceae Sutterella Enterobacteriaceae Parasutterella Ruminococcaceae Lachnospiraceae Escherichia/Shigella Veillonellaceae Faecalibacterium Others Oscillibacter Lachnospiracea_incertae_sedis Dialister Megamonas Others 10
연계서비스 mrna Profiling Service (RNA-Seq+qPCR Array Service) Custom DNA Sequencing Service (Sanger Sequencing) mrna Profiling Service (RNA-Seq + qpcr Array Service) RNA-seq 과 qpcr Array System 은숙련된전문가에의하여수행되는고품질서비스입니다. RNA Sequencing Service 를통해샘플간발현값의차이를확인할수있습니다. 풍부한 Bioinformatics 인프라를갖추고있어차별화된분석패키지 (Gene Expression, DEG, GO 등 ) 를제공합니다. qpcr Array Service 는 Real-Time PCR 전문가들의지침서인 *MIQE Guidline 을준수하여논문에게재가능한정확한 Real-Time PCR 데이터를신속하게제공합니다. *Bustin, S.A., et al. 2009. The MIQE Guidelines: Minimum Information for Publication of Quantitative Real-Time PCR Experiments, Clinical Chemistry 55:4, 611-622. 주요특징 RNA-seq 및 qpcr array service 로유전자탐색및검증까지한번에해결! RNA Sequencing Total RNA Sequencing mrna Sequencing Small RNA Sequencing Gene Expression Analysis Service qpcr Array Service Primer Optimization Service qpcr Array System: Customized qpcr panel kit Single Gene qpcr Primer Set 서비스과정 이메일주문접수 샘플발송 / 회수 RNA 추출및 QC Library Prep. NGS Data 제공 qpcr Array Service 샘플발송 / 회수 RNA 추출및 QC cdna 합성 qpcr 및데이터분석 Data 제공 서비스문의및상담 RNA-seq 관련이메일 : NGS_support@bioneer.co.kr, 전화번호 : 042-930-8553 qpcr Array Service 관련이메일 : qpcrarray@bioneer.com, 전화번호 : 042-930-8673 11
Custom DNA Sequencing Service (Sanger Sequencing) DNA/RNA 추출부터 primer design 을포함하여 PCR optimization 및 sequencing 까지일련의과정에대한맞춤서비스입니다. 주요특징 염기서열분석을위한 Total Solution 제공바이오니아의다양한추출 Application 고객 Needs의맞춤디자인자동분석장비 ABI3730XL을이용하여신뢰도높은분석결과제공 서비스과정 G A T AA A T C T GG T C TT A TT CC 온라인주문접수샘플회수 Sequencing Data 제공 서비스문의및상담 이메일 : sequencing@bioneer.co.kr, 전화번호 : 042-930-8550, 8553 12
Next Generation Sequencing Service Sample Submission Guidelines Sample Requirements DNA RNA FFPE Quantity 1 µg 1 µg Concentration 50 ng/µl 65 ng/µl Purity OD 260/280 = 1.5~2.2 RIN 7.0 Condition Without degradation / RNA contamination Without degradation / DNA contamination Surface area 25 mm 2 Sample depth 40 microns Sample volume 1 mm 3 Nucleated cellularity 80% Tumor content 20% Sample Package 샘플은 1.5 ml microcentrifuge tube 에담아 sealing 후샘플팩에넣어발송합니다. 배송중 degradation 이되지않도록충분한드라이아이스를동봉하며, 파손에유의하여완충재를넣어발송합니다. ( 배송지연에의한샘플변성방지를위해금요일또는공휴일전날배송은삼가주시기바랍니다.) Contact Us 보내는주소 : 대전광역시대덕구문평서로 8-11 바이오니아 Sequencing 팀연락처 : 042-930-8575, NGS_support@bioneer.co.kr 상담시간 : 평일오전 9:00 ~ 오후 6:00 ( 주말, 휴일휴무 ) 13
Sequencing Platform HiSeq 2500 HiSeq 4000 NovaSeq 6000 MiSeq HiSeq X Ion Proton 14
www.bioneer.co.kr Contact Us Bioneer Corporation 8-11 Munpyeongseo-ro, Daedeok-gu Daejeon, 34302, Republic of Korea Tel: +82-42-930-8777 (Korea: 1588-9788) Fax: +82-42-930-8688 E-mail: sales@bioneer.com Bioneer Inc. 155 Filbert St. Suite 216 Oakland, CA 94607, USA Toll Free: +1-877-264-4300 Fax: +1-510-865-0350 E-mail: order.usa@bioneer.us.com Bioneer R&D Center Korea Bio Park BLDG #B-702 700 Daewangpangyo-ro, Bundang-gu, Seongnam-si Gyeonggi-do, 13488, Republic of Korea Tel: +82-31-628-0500 Fax: +82-31-628-0555 Copyright 2019 by Bioneer Corporation. All Rights Reserved.