GSM5592350: rna_P1_10_STR_GAATTCGT-GTACTGAC [P1_STR_10]; Mus musculus; RNA-Seq
Freshly dissected tissue was flash-frozen in liquid nitrogen and frozen at -80° C until RNA could be extracted. Total RNA was disrupted in Trizol (Life Technologies, Carlsbad, CA, USA) with a mechanical pestle according manufacturer's specifications. The aqueous phase was precipitated in isopropanol, resuspended in nuclease-free water, treated with DNase I (New England Biolabs, Ipswich, MA, USA), and cleaned up using a Zymo RNA Clean & Concentrator™-25 kit (Zymo Research, Irvine, CA, USA) according to manufacturer's specifications. Prior to library preparation, total RNA was checked for purity using a NanoDrop ND-1000 spectrophotometer, integrity using RNA Nano chips on an Agilent 2100 Bioanalyzer, and concentration using a Qubit fluormeter. Illumina sequencing libraries were generated with the TruSeq Stranded mRNA HT kit (Illumina) using an Eppendorf ePMotion 5075 robot, and were sequenced to a depth 45-60 million reads per sample on Hi-Seq 2500 instruments at the Roy J. Carver Biotechnology Center at the University of Illinois.