alias: MB_rep2
title: RNA-seq of Proliferating myoblast( C2C12 mouse)
design: The cell number was calculated using Countess TM automated cell counter (Life Technologies) using 3 serial dilutions of cell suspension. RNA was isolated using Qiagen RNA isolation kit and quantified as per supplierÕs protocol. The quality of RNA was verified using BioAnalyser. Mean cell number was used as a reference to draw RNA proportional to one million cells. 3 _l of 1:10 dilution ERCC spike-in RNA was added. The spiked total cellular RNA was DNase treated to remove trace amounts of DNA (DNA-freeTM kit, AmbionTM Cat. No AM1906) and 4 _g of DNase-treated spiked RNA was processed to remove ribosomal RNA (RiboMinusTM Eukaryotic kit V2, AmbionTM cat no A15020). The ribominus RNA was quality checked using BioAnalyser. Finally, the purified mRNA was used for cDNA synthesis and library preparation using NEBNext¨ UltraTM Directional RNA Library Prep Kit for Illumina¨ (New England Biolabs¨ cat. No. E7420). The library thus prepared was quality checked on BioAnalyser and  paired-end sequencing was carried out.
identifiers.namespace: SUB1909195
identifiers.id: MB_rep2