Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
1993429 | Methods | 2014 | 9 Pages |
•BRIC-seq method is suitable for the determination of genome-wide RNA stability.•BRIC-seq can determine the RNA half-life of each transcript.•Protocols for BrU-labelling of RNA, cDNA library construction and data analysis.
We recently developed a novel transcriptome analysis method, termed 5′-bromo-uridine (BrU) immunoprecipitation chase-deep sequencing analysis (BRIC-seq). BRIC-seq enables the determination of genome-wide RNA stability by chasing chronological decreases of BrU-labeled RNAs under physiologically undisturbed conditions. The RNA half-life of each transcript is calculated from the decreasing number of BrU-labeled RNA sequence tags measured by deep sequencing of BrU-labeled RNAs. Here, we describe a detailed protocol and provide tips for BRIC-seq, followed by computational analysis.