GSM5501288: Total RNA from primary myoblasts CP2; Homo sapiens; RNA-Seq
Total RNA was isolated and purified from 70-80% confluent myoblast cell culture preparations (passage 4) using Trizol reagent (Ambion - Life Technologies) and a Qiagen RNeasy Mini Kit according to manufacturer's recommendations. Total RNA examples were checked for quality using RINs generated from Agilent Bioanalyzer 2100. RNA quantity was determined using a Qubit fluorometer. The Illumina TruSeq Stranded mRNA Library Preparation Kit was used to prepare sequencing libraries from 1 ug of high-quality RNA samples (RIN>7). The Kit procedure was performed without modifications. This procedure included mRNA purification and fragmentation, cDNA synthesis, 3' end adenylation, Illumina adapter ligation, library PCR amplification and validation. An IlluminaIllumina  NextSeq 500 sequencer was used to sequence the libraries with the production of single-end, 75 bp reads at the depth of 20-25 M reads per sample.