GSM9690800: Control sample 2 C2C12 cells, non-targeting shRNA; Mus musculus; RNA-Seq
Forty-eight hours after transfection, cells were lysed with QIAzol® and total RNA was extracted using the PureLink RNA Mini Kit. Library preparation and RNA sequencing were performed at the McGill University Genome Centre. cDNA synthesis was carried out using the NEBNext RNA First Strand Synthesis Module and the NEBNext Ultra Directional RNA Second Strand Synthesis Module (New England BioLabs). Subsequent library preparation steps were completed using the NEBNext Ultra II DNA Library Prep Kit for Illumina. Libraries were quantified using the Kapa Illumina GA with Revised Primers-SYBR Fast Universal kit, and fragment size distribution was assessed using a LabChip GX instrument. Libraries were normalized, pooled, denatured in 0.05 N NaOH, neutralized using HT1 buffer, and loaded at 225 pM onto an Illumina NovaSeq S4 lane using the Xp workflow. A 1% PhiX control library was included.