title: GSM6426647: mdmEDL_replicate2; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA from the collected muscles was extracted according to the recommended protocol in the Qiagen Fibrous Tissue Total RNA extraction mini kit. RNA concentration was measured using a Qubit RNA Broad-Range assay, and RNA integrity number (RIN) was assessed with Agilent 2100 Bioanalyzer RNA 6000 Nano assay. Samples with a concentration >20ng/ul, RIN>7 were chosen for the subsequent cDNA library preparation. cDNA libraries from the selected samples were prepared with Illumina TruSeq Stranded Total RNA Library Prep Kit. Quality of the cDNA libraries was assessed with KAPA Library Quantification qPCR Kit for Illumina sequencing platforms, and library fragment sizes were tested using an Agilent 2100 Bioanalyzer High Sensitivity dsDNA quantification assay. The mean library sizes of the samples were between 256-323bp. Prepared cDNA libraries were sequenced in an Illumina NextSeq 500 high throughput sequencer following sequencing guidelines recommended by Illumina. Short 75 bp paired-end reads were generated from each library, and the samples were sequenced using a total of five sequencing runs. The sequenced library coverage varies between 9.5 - 100 million reads with a median of 23 million reads.