通过下一代测序进行小 RNA 分析的条形码 cDNA 文库制备。
Barcoded cDNA library preparation for small RNA profiling by next-generation sequencing.
机构信息
Laboratory of RNA Molecular Biology, Howard Hughes Medical Institute, The Rockefeller University, 1230 York Avenue, Box 186, NY 10065, USA.
出版信息
Methods. 2012 Oct;58(2):164-70. doi: 10.1016/j.ymeth.2012.07.030. Epub 2012 Aug 7.
Abstract
The characterization of post-transcriptional gene regulation by small regulatory (20-30 nt) RNAs, particularly miRNAs and piRNAs, has become a major focus of research in recent years. A prerequisite for characterizing small RNAs is their identification and quantification across different developmental stages, and in normal and disease tissues, as well as model cell lines. Here we present a step-by-step protocol for generating barcoded small RNA cDNA libraries compatible with Illumina HiSeq sequencing, thereby facilitating miRNA and other small RNA profiling of large sample collections.
摘要
近年来，小调控 RNA（20-30nt），特别是 miRNA 和 piRNA，在后转录基因调控的特性研究方面成为了主要关注点。鉴定和定量分析不同发育阶段、正常组织和疾病组织以及模型细胞系中的小 RNA，是其特性研究的前提条件。本研究提供了一种分步协议，用于生成兼容 Illumina HiSeq 测序的带条码的小 RNA cDNA 文库，从而促进了 miRNA 和其他小 RNA 对大样本集的分析。
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