GSM1166239: Rapa_S8; Mus musculus; RNA-Seq
Whole frozen heart tissue was crushed to a powder in liquid nitrogen. The powdered tissue was then treated with Qiasol lysis buffer and extracted for total RNA on a Qiacube robot using the RNeasy mini kit (Invitrogen). The extracted RNA was quantitated using a NanoDrop and RNA quality was measured via BioAnalyzer chip (Agilent). Oligo-dT purification of polyadenylated RNA, and reverse transcription to create cDNA. The cDNA is fragmented, blunt-ended, and ligated to indexed (barcoded) adaptors. The library is size selected (in the case of paired-end RNA-Seq), size distribution validated using capillary electrophoresis, and quantified using fluorimetry (PicoGreen) and via Q-PCR.