CLIP: Construction of cDNA libraries for high-throughput sequencing from RNAs cross-linked to proteins in vivo

被引：65

作者：

Wang, Zhen ^{[1
]}

Tollervey, James ^{[1
]}

Briese, Michael ^{[1
]}

Turner, Daniel ^{[2
]}

Ule, Jernej ^{[1
]}

机构：

[1] MRC, Mol Biol Lab, Cambridge CB2 0QH, England

[2] Wellcome Trust Sanger Inst, Cambridge CB10 1SA, England

来源：

METHODS | 2009年 / 48卷 / 03期

基金：

欧洲研究理事会;

关键词：

CLIP; UV cross-linking; Immunoprecipitation; RNA-protein binding; High-throughput sequencing; TARGETS; BRAIN; CELLS;

D O I：

10.1016/j.ymeth.2009.02.021

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

UV cross-linking and immunoprecipitation assay (CLIP) can identify direct interaction sites between RNA-binding proteins and RNAs in vivo, and has been used to study several proteins in tissues and cell cultures. The main challenge of the method is to specifically amplify the low amount of isolated RNA. The current protocol is optimised for efficient RNA purification and ligation of barcoded RNA adapters, High-throughput sequencing of the multiplexed cDNA library allows for a comprehensive coverage of the target sequences. (C) 2009 Elsevier Inc. All rights reserved.

引用

页码：287 / 293

页数：7

共 12 条

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