RNA-seq of mus musculus: mouse liver tissue
Total RNA was isolated from mouse liver tissue using TRIzol reagent (Invitrogen, USA) according to the manufacturers protocol. In this study, we used an m6A-specific antibody (Sigma-Aldrich, ABE572) for immunoprecipitation RNA. Briefly, poly (A) RNA was captured by VAHTS 2X Frag/Prime Buffer. Then one part of the RNA fragment was used to construct the RNA-seq library, and the other part was used for m6A RNA immunoprecipitation through the GenSeqTM m6A-MERIP kit (GenSeq Inc., Cyberjaya, Malaysia), which was used to construct the m6A-seq library. All operations were carried out in accordance with the manufacturer's instructions. RNA input samples without immunoprecipitation and m6A input samples were used for the generation of RNA-seq libraries. The library quality was evaluated with a Bioptic Qsep100 Analyzer (Bioptic lnc., Taiwan, China). Library sequencing was performed on an Illumina NovaSeq instrument with 150 bp paired-end reads.