GSM8502666: Bisulfite-seq_WT1; Mus musculus; Bisulfite-Seq
Total RNAs were isolated by TRIzol reagent from 40mg tissues after homogenization with a dounce homogenizer. Poly-A tailed RNAs were enriched by a GenElute mRNA Miniprep Kit (Sigma-Aldrich). Treated mRNAs with 0.5% unmethylated RNA controls in vitro transcribed from lambda genome were treated using EZ RNA Methylation Kit (Zymo Research), with the following modifications: 500 ng of mRNAs were bisulfited using two cycles of 10 min denaturation at 70 °C followed by 45 min at 54 °C. Strand-specific libraries were constructed with VAHTS Stranded mRNA-seq Library Prep Kit for Illumina (vazyme), and sequenced using NovaSeq 6000 (Illumina).