Direct sequencing of small genomes on the Pacific Biosciences RS without library preparation.

Direct sequencing of small genomes on the Pacific Biosciences RS without library preparation.
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DOI:
10.2144/000113962
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发表时间:
2012-12
期刊:
影响因子:
2.7
通讯作者:
Swerdlow H
Swerdlow H
中科院分区:
工程技术4区
文献类型:
--
作者:
Coupland P;Chandra T;Quail M;Reik W;Swerdlow H

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我们在 Pacific Biosciences RS 测序仪 (PacBio) 上开发了一种针对小 DNA 分子的测序方法,无需标准文库制备。迄今为止,这种方法已应用于单链和双链病毒基因组、细菌质粒、用于 DNA 修饰分析的质粒载体模型以及覆盖整个细菌基因组的线性 DNA 片段的测序。使用直接测序可以从少至 1 ng 的 DNA 生成序列数据,与当前的方案相比具有显着优势,目前的方案通常需要 400-500 ng 的剪切 DNA 来进行文库制备。
We have developed a sequencing method on the Pacific Biosciences RS sequencer (the PacBio) for small DNA molecules that avoids the need for a standard library preparation. To date this approach has been applied toward sequencing single-stranded and double-stranded viral genomes, bacterial plasmids, plasmid vector models for DNA-modification analysis, and linear DNA fragments covering an entire bacterial genome. Using direct sequencing it is possible to generate sequence data from as little as 1 ng of DNA, offering a significant advantage over current protocols which typically require 400–500 ng of sheared DNA for the library preparation.
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