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使用多重置换扩增的单细胞基因组测序。

Single-cell genomic sequencing using Multiple Displacement Amplification.

作者信息

Lasken Roger S

机构信息

J. Craig Venter Institute, 11149 N. Torrey Pines Rd., Suite 200, La Jolla, CA 92037, USA.

出版信息

Curr Opin Microbiol. 2007 Oct;10(5):510-6. doi: 10.1016/j.mib.2007.08.005. Epub 2007 Oct 17.

Abstract

Single microbial cells can now be sequenced using DNA amplified by the Multiple Displacement Amplification (MDA) reaction. The few femtograms of DNA in a bacterium are amplified into micrograms of high molecular weight DNA suitable for DNA library construction and Sanger sequencing. The MDA-generated DNA also performs well when used directly as template for pyrosequencing by the 454 Life Sciences method. While MDA from single cells loses some of the genomic sequence, this approach will greatly accelerate the pace of sequencing from uncultured microbes. The genetically linked sequences from single cells are also a powerful tool to be used in guiding genomic assembly of shotgun sequences of multiple organisms from environmental DNA extracts (metagenomic sequences).

摘要

现在可以使用通过多重置换扩增(MDA)反应扩增的DNA对单个微生物细胞进行测序。细菌中仅有的几飞克DNA被扩增成适合用于构建DNA文库和桑格测序的微克级高分子量DNA。当通过454生命科学方法直接用作焦磷酸测序的模板时,MDA生成的DNA也表现良好。虽然单细胞的MDA会丢失一些基因组序列,但这种方法将大大加快未培养微生物的测序速度。来自单细胞的遗传连锁序列也是一种强大的工具,可用于指导从环境DNA提取物(宏基因组序列)中对多种生物的鸟枪法序列进行基因组组装。

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