GSM1782908: mouse embronic stem cells IP replicate A; Mus musculus; OTHER
Total RNA of the indicated cell types was extracted using 5PRIME RNA purification kit. Where indicated mRNA enriched RNA was purified by oligo dT selection. Fragmentation was carried out using Ambion fragmentation reagents in 70ºc.Competitive elution was carried out using free m1A nucleoside  (330 microMolar m1A in 10 mM Tris pH 7.4 ,375 mM NaCl 0.1% NP-40. Immunoprecipitation was done using MBL anti-m1A antibody D345-3 mAb.Dimmroth rearrangement was performed by heating the samples to either 50ºC or 60ºC for 1H under pH 10.4 conditions. Libraries were prepared according to NEB's instructions accompanying the NEBNext® Ultra™ RNA Library Prep Kit for Illumina (Cat# 7530).