GSM2454763: P12_m6A_Null_IP rep 2; Mus musculus; RNA-Seq
Total RNA was isolated using TriZol. Total RNA was subjected to two rounds of poly(A) RNA selection, using the MicroPoly(A) Purist kit, (Life Technologies) according to the manufacturer's protocol. RNA was fragmented by incubating for 45 second with pre-warmed Zinc chloride buffer (10 mM ZnCl2, 10mM Tris-HCl, ph 7.0), to fragment the RNA to ~100 nucleotide long fragments. The reaction was stopped with 0.2M EDTA and immediately placed on ice. A fraction of teh RNA was subjected to immunoprecipitation with a anti-m6A antibody. A strand specific protocol was used to generate cDNA from input and anti-m6A immunoprecipitated samples.