GSM6336619: +MyoD+F/R/C+CP_3; Mus musculus; RNA-Seq
Total RNA was isolated using the RNAeasy  Mini Kit (Qiagen, #74104) as per the manufacturer's protocol. Samples with RIN above 8 were only used for further library preparation steps and library preparation was conducted according to the TruSeq Stranded mRNA protocol (Illumina, Inc., California, USA). Briefly, poly-A enrichment was carried out for total RNA (100-1000 ng), followed by reverse-transcription of poly-A enriched RNA into cDNA. The cDNA was then ligated with TrueSeq adapters including unique dual indices (UDI) after fragmentation, end-repair and adenylation steps. Using PCR, cDNA containing TruSeq adapters on both ends was selectively amplified which was further applied to a Fragment Analyzer to determine the quality and quantity of libraries. The average fragment size was approximately 360 bp. 