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Single-molecule DNA sequencing of a viral genome

peacock 添加于 2009-8-24 10:22 | 2055 次阅读 | 0 个评论

作者
Harris TD, Buzby PR, Babcock H, Beer E, Bowers J, Braslavsky I, Causey M, Colonell J, Dimeo J, Efcavitch JW, Giladi E, Gill J, Healy J, Jarosz M, Lapen D, Moulton K, Quake SR, Steinmann K, Thayer E, Tyurina A, Ward R, Weiss H, Xie Z
摘要
The full promise of human genomics will be realized only when the genomes of thousands of individuals can be sequenced for comparative analysis. A reference sequence enables the use of short read length. We report an amplification-free method for determining the nucleotide sequence of more than 280,000 individual DNA molecules simultaneously. A DNA polymerase adds labeled nucleotides to surface-immobilized primer-template duplexes in stepwise fashion, and the asynchronous growth of individual DNA molecules was monitored by fluorescence imaging. Read lengths of >25 bases and equivalent phred software program quality scores approaching 30 were achieved. We used this method to sequence the M13 virus to an average depth of >150x and with 100% coverage; thus, we resequenced the M13 genome with high-sensitivity mutation detection. This demonstrates a strategy for high-throughput low-cost resequencing.
详细资料
- 关键词: Algorithms; Bacteriophage M13/*genetics; Computational Biology/methods; DNA Primers; DNA, Viral/chemistry/*genetics; *Genome, Viral; Mutation; Sequence Alignment; Sequence Analysis, DNA/*methods; Software; Templates, Genetic
- 文献种类:期刊
- 期刊名称: Science (New York, N.Y.)
- 期刊缩写: Science
- 期卷页: 2008年第320卷第5872期 106-109页
- 地址: Helicos BioSciences Corporation, One Kendall Square, Cambridge, MA 02139, USA. tharris@helicosbio.com
- ISBN: 1095-9203
- 备注:PMID:18388294
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测序技术单细胞
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