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一种用于下一代测序样本制备的微生物组DNA富集方法。

A Microbiome DNA Enrichment Method for Next-Generation Sequencing Sample Preparation.

作者信息

Yigit Erbay, Feehery George R, Langhorst Bradley W, Stewart Fiona J, Dimalanta Eileen T, Pradhan Sriharsa, Slatko Barton, Gardner Andrew F, McFarland James, Sumner Christine, Davis Theodore B

机构信息

New England Biolabs, Ipswich, Massachusetts.

出版信息

Curr Protoc Mol Biol. 2016 Jul 1;115:7.26.1-7.26.14. doi: 10.1002/cpmb.12.

Abstract

"Microbiome" is used to describe the communities of microorganisms and their genes in a particular environment, including communities in association with a eukaryotic host or part of a host. One challenge in microbiome analysis concerns the presence of host DNA in samples. Removal of host DNA before sequencing results in greater sequence depth of the intended microbiome target population. This unit describes a novel method of microbial DNA enrichment in which methylated host DNA such as human genomic DNA is selectively bound and separated from microbial DNA before next-generation sequencing (NGS) library construction. This microbiome enrichment technique yields a higher fraction of microbial sequencing reads and improved read quality resulting in a reduced cost of downstream data generation and analysis. © 2016 by John Wiley & Sons, Inc.

摘要

“微生物组”用于描述特定环境中微生物群落及其基因,包括与真核宿主或宿主部分相关的群落。微生物组分析中的一个挑战涉及样本中宿主DNA的存在。在测序前去除宿主DNA可使目标微生物组群体的序列深度更大。本单元描述了一种新的微生物DNA富集方法,即在构建下一代测序(NGS)文库之前,将甲基化的宿主DNA(如人类基因组DNA)与微生物DNA选择性结合并分离。这种微生物组富集技术可产生更高比例的微生物测序读数并提高读数质量,从而降低下游数据生成和分析的成本。©2016约翰威立国际出版公司。

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