m6A-Seq and RNA-Seq analyses of female and male mice livers
500ng mRNA (100ng each mouse, n=5) purified with Oligo(dT) Dynabeads from total RNA isolated with TRIzol. 5g of fragmented mRNA was incubated with 12g of anti-m6A antibody. On average, 5g of purified mRNA yielded tens of nanograms of immunopreciptated mRNA fragments.  Input and immunoprecipitated RNA fragments were subjected to first-strand cDNA synthesis and library prep using TruSeq stranded mRNA library prep kit (Illumina) and sequenced on an Illumina HiSeq 3000 with single-end 50-bp read length.
PTTSmm_6A.In.HF4