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PMID: 19272451 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

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

Methods (San Diego, Calif.) ·Vol. 48 ·No. 3 ·2009-07-00 ·Pages 287-93

Wang Z, Tollervey J, Briese M, Turner D, Ule J

Abstract

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.

MeSH Terms
Animals Cells, Cultured Cross-Linking Reagents/chemistry Gene Expression Profiling/methods Gene Library Immunoprecipitation Mice RNA/genetics RNA-Binding Proteins/genetics Sequence Analysis/methods Ultraviolet Rays
Chemicals
Cross-Linking Reagents RNA-Binding Proteins RNA
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Wang Zhen
MRC-Laboratory of Molecular Biology, Hills Road, Cambridge CB20QH, UK.
Tollervey James
Briese Michael
Turner Daniel
Ule Jernej
Article Info
Journal
Methods (San Diego, Calif.)
Abbr.
Methods
ISSN
1095-9130
Published
2009-07-00
Epub
2009-00-09
Pages
287-93
Language
English
Region
United States
NLM ID
9426302
Subset
IM
Grants
Wellcome Trust · 089701 · United Kingdom
Medical Research Council · MC_U105185858 · United Kingdom
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