GSM9131528: Old CTR rep2; Mus musculus; RNA-Seq
Satellite cells from control and AR(i)sat-/Y mice were isolated by FACS as described (doi:10.3791/65215). For RNA isolation from FACS-isolated MuSC, the RNeasy Plus Micro Kit (Qiagen, 74034) was employed according to the manufacturer's instructions P3388 : Full length cDNA were generated from 250 pg to 2 ng of total RNA using SMART-Seq® mRNA Kit (Takara Bio Europe, Saint Germain en Laye, France) according to manufacturer's instructions with 12 cycles of PCR for cDNA amplification by Seq-Amp polymerase. Six hundreds pg of pre-amplified cDNA were then used as input for Tn5 transposon tagmentation followed by 12 cycles of library amplification using Nextera XT DNA Library Preparation Kit and IDT for Illumina DNA/RNA UD Indexes, Tagmentation (Illumina, San Diego, USA). Following purification with SPRIselect beads (Beckman-Coulter, Villepinte, France), the size and concentration of libraries were assessed using Bioanalyzer 2100 system (Agilent technologies, Les Ulis, France).