- m6A-label-seq: A metabolic labeling protocol to detect transcriptome-wide mRNA N 6-methyladenosine (m6A) at base resolution.
m6A-label-seq: A metabolic labeling protocol to detect transcriptome-wide mRNA N 6-methyladenosine (m6A) at base resolution.
STAR protocols (2022-01-22)
Xiao Shu, Jie Cao, Jianzhao Liu
PMID35059657
摘要
We describe here a metabolic labeling protocol for detecting cellular transcriptome-wide mRNA N 6 -methyladenosine (m6A) at base resolution. By feeding cells an analog of methionine, potential mRNA m6A forming sites are replaced with the N 6 -allyladenosine (a6A). A mild chemical iodination of a6A in RNA results in its opposite base misincorporation during RNA reverse transcription, and thus m6A locations could be precisely identified in the high-throughput sequencing. m6A-label-seq provides a strategy to label and identify cellular epitranscriptomic modification sites. For complete details on the use and execution of this profile, please refer to Shu et al., 2020.
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