alias: E-MTAB-5846:S12_p
title: NextSeq 500 paired end sequencing; Enhanced energetic state and protection from oxidative stress in human myoblasts overexpressing BMI1
design: Enhanced energetic state and protection from oxidative stress in human myoblasts overexpressing BMI1
library_construction_protocol: Immortalised human myoblasts (isolated from DMD and histologically normal muscle biopsies) were obtained from the MRC Centre for Neuromuscular Diseases Biobank (38) and used in this study under the same ethics as above. Cells were maintained in Skeletal Muscle Medium plus supplements (Promocell Kit, C-23060), 10% FBS (PAA A15-151), 1X Glutamax (Invitrogen 35050038), 5ug/ml Gentamicin (Sigma G1272). Differentiation was induced at confluence by replacing the growth medium with DMEM, 2% HS. Three independently prepared normal and DMD myoblasts cell cultures infected with GFP and BMI1Over lentiviral particles and induced to differentiate for 2 days  RNA extraction was carried out using the RNeasy Micro purification kit (Qiagen) and DNase digestion was performed to remove genomic DNA. cDNA synthesis was performed with SuperScript III Reverse Transcriptase Kit (Invitrogen).  One microgram of total RNA was used for library preparation, according to the Illumina TruSeq mRNA sample preparation protocol (Illumina, San Diego),
identifiers.namespace: QMUL
identifiers.id: E-MTAB-5846:S12_p
identifiers.namespace: Queen Mary University of London
identifiers.id: E-MTAB-5846:S12_p