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核糖体RNA去除以高效利用RNA测序能力。

Ribosomal RNA depletion for efficient use of RNA-seq capacity.

作者信息

O'Neil Dominic, Glowatz Heike, Schlumpberger Martin

机构信息

Qiagen, Hilden, Germany.

出版信息

Curr Protoc Mol Biol. 2013 Jul;Chapter 4:Unit 4.19. doi: 10.1002/0471142727.mb0419s103.

Abstract

Ribosomal RNA (rRNA) is the most highly abundant component of RNA, comprising the majority (>80% to 90%) of the molecules present in a total RNA sample. Depletion of this rRNA fraction is desirable prior to performing an RNA-seq reaction, so that sequencing capacity can be focused on more informative parts of the transcriptome. This unit describes an rRNA depletion method based on selective hybridization of oligonucleotides to rRNA, recognition with a hybrid-specific antibody, and removal of the antibody-hybrid complex on magnetic beads.

摘要

核糖体RNA(rRNA)是RNA中含量最为丰富的成分,在总RNA样本中占分子总数的大部分(>80%至90%)。在进行RNA测序反应之前,去除这部分rRNA是很有必要的,这样测序能力就可以集中在转录组中更具信息性的部分。本单元描述了一种基于寡核苷酸与rRNA选择性杂交、用杂交特异性抗体识别以及在磁珠上去除抗体 - 杂交复合物的rRNA去除方法。

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