GSM8008572: WT_PRED2; Mus musculus; RNA-Seq
RNA-seq was conducted on RNA extracted from myocardial tissue. Total RNA was extracted from cryo-pulverized, atria- and blood-deprived hearts with Trizol (#15596026; Thermo Fisher Scientific, Waltham, MA) and re-purified using the RNeasy Mini Kit (Cat #74104; Qiagen, Germantown, MD). RNA-seq was performed at the DNA Core (CCHMC) blinded to treatment groups. 150 to 300 ng of total RNA determined by Qubit (Cat#Q33238; Invitro-gen, Waltham, MA) high-sensitivity spectrofluorometric measurement was poly-A selected and reverse tran-scribed using Illumina's TruSeq stranded mRNA library preparation kit (Cat# 20020595; Illumina, San Diego, CA). Each sample was fitted with one of 96 adapters containing a different 8 base molecular barcode for high level multiplexing. After 15 cycles of PCR amplification, libraries were sequenced.