大规模基因组测序的原理与方法培训.ppt

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2018-02-13 发布于天津
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大规模基因组测序的原理与方法培训.ppt

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Consensus Sequence Gap Mis-Assembly (Inverted) Shotgun Sequencing III: FINISHING Consensus Mis-Assembly (Inverted) Shotgun Sequencing III: FINISHING Shotgun Sequencing III: FINISHING High Accuracy Sequence: 1 error/ 10,000 bases Consed软件显示序列组装结果界面 1、Filling “intraclone gaps” BAC-----453F3’s finishing Sp6 T7 ? First 4 primers 1 2 3 4 All the contigs walked hundreds’ bps toward the gaps. 453F3’s 2600 reads 12 contigs Overlapping BAC-454F24’s 200 reads + Sp6 T7 1 3 2 4 a b c Second 3 primers 1200bp’s AT-rich , (CATATATA)n repeat. Finally, filled by using ET sequencing Kit . 1240bp’s GC-rich, GC-content is 69.03% ; the BAC’s is 39.98%. We used dGTP Kit filling it. Sp6 T7 Completed sequence ? Sequenced clone BAC selected by end-sequence 113L10 324K11 173F11 101A4 167P17 586C2 116K5 572B2 2544N5 R-155E14 2006P23 2306M15 R-149E15 60K ? Gap filling by end sequences 2、Filling “interclone gaps” The actual and predicted fingerprint of R-260J13 digested with HindIII Lane 1: marker, Lane 2: R-260J13 digested with HindIII, 3 : the predicted 克隆211B19组装后的序列的错误率为零 Whole Genome Shotgun This bacterium has a circular genome structure with 2,689,445 base pairs, the second largest one of thermophiles decoded completely to date. Circular representation of the genome of T. tengcongensis What is under heaven is for all. Sun Yat-sen, the father of modern China 天下为公 /rice DDBJ/EMBL/GenBank:AAA国际一流测序生产线 7万克隆，3000万碱基/天高产出、低成本： $/bp?￥/bp?美分/bp?分/bp 基因组学：数据导向的大科学有数据才是硬道理世上无难事只要肯登攀 Contigs:127,550 (N50=6,688 bp) Scaffolds: 102,444 (N50=11,764 bp) Quality: 546 bp at Q20 De Novo Sequencing the Genome in BIG Hu Songnian Beijing Institute of Genomics, Chinese Academy of Sciences Next Generation Sequencing (NGS) Technology Second generation sequencers 454 1 Solexa 3 SOLiD 5 De novo sequencing RNA-seq, Re-sequencing ChIP-seq， Meth-seq Metagenomics