Transcriptome-Wide Mapping of m6 A and m6 Am at Single-Nucleotide Resolution Using miCLIP.
Curr Protoc Mol Biol
; 126(1): e88, 2019 04.
Article
em En
| MEDLINE
| ID: mdl-30874375
ABSTRACT
The most prevalent modified base in mRNA, N6 -methyladenosine (m6 A), is found in several thousand transcripts, typically near the stop codon, although it can occur anywhere in the mRNA. In addition, the highly similar nucleotide N6 ,2'-O-dimethyladenosine (m6 Am), which is difficult to distinguish from m6 A, occurs as the first transcribed nucleotide of certain transcripts. Both the m6 A and m6 Am modifications have been implicated in numerous biological processes, and their precise mapping is crucial to understanding their functions. To address this need, we developed miCLIP, a method that maps both m6 A and m6 Am at individual nucleotide resolution. Here we describe the miCLIP protocol, with slight improvements to the initially published protocol for both the experimental methodology and bioinformatics analysis. © 2019 by John Wiley & Sons, Inc.
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01-internacional
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MEDLINE
Idioma:
En
Ano de publicação:
2019
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Article