使用“单病毒基因组学”对单个病毒粒子进行分离和基因组分析。
Isolation and genome analysis of single virions using 'single virus genomics'.
作者信息
Zeigler Allen Lisa, Ishoey Thomas, Novotny Mark A, McLean Jeffrey S, Lasken Roger S, Williamson Shannon J
机构信息
Department of Microbial and Environmental Genomics, The J. Craig Venter Institute, USA.
出版信息
J Vis Exp. 2013 May 26(75):e3899. doi: 10.3791/3899.
Whole genome amplification and sequencing of single microbial cells enables genomic characterization without the need of cultivation (1-3). Viruses, which are ubiquitous and the most numerous entities on our planet (4) and important in all environments (5), have yet to be revealed via similar approaches. Here we describe an approach for isolating and characterizing the genomes of single virions called 'Single Virus Genomics' (SVG). SVG utilizes flow cytometry to isolate individual viruses and whole genome amplification to obtain high molecular weight genomic DNA (gDNA) that can be used in subsequent sequencing reactions.
单个微生物细胞的全基因组扩增和测序能够在无需培养的情况下实现基因组特征分析(1-3)。病毒无处不在，是地球上数量最多的实体(4)，且在所有环境中都很重要(5)，但尚未通过类似方法得以揭示。在此，我们描述了一种用于分离和鉴定单个病毒粒子基因组的方法，称为“单病毒基因组学”(SVG)。SVG利用流式细胞术分离单个病毒，并通过全基因组扩增获得可用于后续测序反应的高分子量基因组DNA(gDNA)。
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