GSM5597512: KO-MI-1; Mus musculus; RNA-Seq
Hearts were harvested, full ventricles (above and below MI ligation) were isolated, and immediately homogenized in Trizol (Invitrogen) according to the manufacturer's protocol. Samples were lysed on ice for 20 minutes, then chloroform was added, and the resulting aqueous RNA phase was isolated. Total RNA was processed using RNeasy Fibrous Tissue MiniKit (Qiagen, #74704). RNA sequencing libraries were prepared using the NEBNext Ultra RNA Library Prep Kit for Illumina using manufacturer's instructions (NEB, Ipswich, MA, USA). Briefly, mRNAs were initially enriched with Oligod(T) beads.  Enriched mRNAs were fragmented for 15 minutes at 94 °C. First strand and second strand cDNA were subsequently synthesized. cDNA fragments were end repaired and adenylated at 3'ends, and universal adapters were ligated to cDNA fragments, followed by index addition and library enrichment by PCR with limited cycles.