GSM4844332: Skeletal Muscle D2-HG male rep2; Mus musculus; RNA-Seq
Skeletal muscle were removed, flash frozen in liquid nitrogen, and RNA was extracted from 20 to 30 mg skeletal  muscle  tissue  using  the QIAGEN  RNeasy  Fibrous  Tissue  Mini  Kit (Qiagen, Hilden, Germany; CAT#74704). RNA concentration was determined using a Qubit 4.0 fluorometer (Life Technologies) and Qubit™ RNA HS Assay Kit (ThermoFisher Scientific, Waltham, MA; CAT#Q32852) according to the manufacturer's instructions. The quality of each RNA sample was checked using an Agilent 4200 Tape Station with Agilent High Sensitivity RNA ScreenTape (Agilent Technologies, Santa Clara, CA, USA; CAT#5067-5579) according to the manufacturer's instructions. For library preparation, poly-A RNA was purified from mRNA using oligo-dT magnetic beads. mRNA was fragmented, and cDNA synthesized. A-tailing was conducted to enable subsequent ligation of paired-end sequencing adapters. Library quality was determined using an Agilent 4200 Tape Station with Agilent High Sensitivity D5000 DNA ScreenTape (CAT#5067-5592, Agilent Technologies, Santa Clara, CA, USA) to ensure correct fragment sizes between 250 and 300 bp. RNA libraries were prepared for sequencing using standard Illumina protocols