title: GSM8217876: A35; Mus musculus; RNA-Seq
library_construction_protocol: RNA was extracted from flash frozen heart tissue with Qiagen RNeasy plus mini kit (74134, Germantown, MD). The integrity of RNA was analyzed at the University of Arizona Sequencing Core with the 2100 Bioanalyzer (Agilent Technologies INC. (serial no.DE24802066; Assay Eukaryote Total RNA Nano Series II.xsy; Chip lot no. WH29BK20) and samples with RINs scores at 8.7-9.5 were sent to Novogene for subsequent sequencing (Sacramento, CA). then converted to library using NEBNext Ultra II RNA Library Prep by Illumina Novogene sequenced each sample according to standard protocol, which entailed 1 ug of RNA for the cDNA library construction using an NEBNext Ultra II RNA Library Prep by Illumina according to the manufacturer's protocol (E7775, New England Biolabs, Ipswich, MA). In brief, mRNA was isolated with the Ribo-Zero Kit (20037135; Illumina, San Diego, CA) then fragmented randomly with a fragmentation buffer. Double-stranded cDNA was synthesized using Il-lumina (RS-122-2001, TrueSeq RNA Sample Preparation, San Diego, CA) and random hexamer primers. The cDNA was then terminally repaired using (AMPure XP system (SPRI bead cleanup); A63881, Beckman Coulter, Indianapolis, IN), 'A' ligated, and added sequencing adaptors. The quality of the enriched 250-350 bp cDNA libraries were assessed with a Qubit 2.0 fluorometer (Thermo Scientific, Waltham, MA, USA), quantitative PCR, and analyzed using an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).