GSM2772595: P0 ovarian total RNA sequencing: YTHDC2 -/-; sample 1; Mus musculus; RNA-Seq
For RNA Bind-n-Seq experiment  randomized 21 nt single-stranded RNAs carrying a central m6A moiety were mixed with Ni-NTA agarose beads alone or bound to 6xHis-SUMO-YTH domain (of hYTHDC2). RNA bound to the beads was phenol chloroform extracted.  For m6a IP experiment testicular RNA was isolated using the Trizol reagent and polyA+ transcripts were purified, fragmented and the fragments were immunopurified using anti-m6A antibody.  Total testicular and ovarian RNAs were isolated using the Trizol reagent. Sequencing libraries  of RNA Bind-n-Seq and m6a IP experiments were prepared using NEBNext® Multiplex Small RNA Library Prep Set for Illumina® (Cat No:E7300) according to manufacturer instructions and sequenced on Illumina 2000 HiSeq platform (EMBL Heidelberg Gene core facility). Sequencing libraries of total testicular and ovarian RNAs were prepared by TruSeq Stranded Total RNA kit with Ribo-Zero Gold and sequenced on the Illumina HiSeq 4000 sequencer (iGE3 Genomics Platform, University of Geneva)