GSM5115391: Inj C3; Mus musculus; RNA-Seq
cultured satellite cells were collected for RNA extraction using Machery Nagel NucleoSpin RNA purification kit Libraries were prepared with TruSeq Stranded Total Library preparation kit according to supplier recommendations. Briefly, the key stages of this protocol are successively, the removal of ribosomal RNA fraction from 400ng of total RNA using the Ribo-Zero Gold Kit; fragmentation using divalent cations under elevated temperature to obtain approximately 300bp pieces; double strand cDNA synthesis using reverse transcriptase and random primers, and finally Illumina adapters ligation and cDNA library amplification by PCR for sequencing. Sequencing was carried out on single-end 75bp of Illumina NextSeq500