从核糖核蛋白（RNP）衍生的调控性非编码RNA生成互补DNA（cDNA）文库。
Generation of cDNA libraries from RNP-derived regulatory noncoding RNAs.
作者信息
Rederstorff Mathieu
机构信息
Biopôle, CNRS UMR 7214 AREMS, Université de Lorraine, Vandoeuvre-lès-Nancy, France.
出版信息
Methods Mol Biol. 2012;925:211-8. doi: 10.1007/978-1-62703-011-3_14.
Next-generation sequencing of noncoding RNA (ncRNA) libraries has become an essential tool for the profiling of ncRNAs and the identification of novel ncRNA species. Here, we describe the generation of a ncRNA-derived complementary DNA (cDNA) library by 3'-tailing of ncRNAs by CTP and poly(A) polymerase, followed by 5'-adapter ligation by T4 RNA ligase and reverse transcription of ncRNAs with an oligo-d(G) anchor primer. Preliminary selection of ncRNAs from ribonucleoprotein particles (RNPs) enables a strong enrichment of the generated libraries with functional regulatory ncRNAs compared to classical approaches.
非编码RNA（ncRNA）文库的下一代测序已成为ncRNA分析和新型ncRNA物种鉴定的重要工具。在此，我们描述了通过CTP和聚（A）聚合酶对ncRNA进行3'加尾，然后通过T4 RNA连接酶进行5'接头连接，并用寡聚-d（G）锚定引物对ncRNA进行逆转录来生成ncRNA衍生的互补DNA（cDNA）文库的方法。与传统方法相比，从核糖核蛋白颗粒（RNP）中初步筛选ncRNA能够使生成的文库强烈富集功能性调节ncRNA。
Zotero 插件