GSM7696408: MyoT-Gtx-N1; Homo sapiens; RNA-Seq
For RNA-seq, 2x105 cells were collected per sample and triplicates were prepared for each condition (MyoB, MyoT-LAM  and MyoT-Gtx) for RNA extraction (Macherey Nagel, 740955) following the manufacturer protocol. Libraries were prepared with TruSeq Stranded Total RNA Sample preparation kit according to supplier recommendations. Briefly, the key stages of this protocol are successively, the removal of ribosomal RNA fraction from 1µg 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.