GSM8878492: O5cornLightB; Mus musculus; RNA-Seq
Total RNA was extracted using Qiagen RNAeasy kit. Total RNA is normalized to 30ng/ul in a total volume of 10ul on the Perkin Elmer Janus Workstation (Perkin Elmer, Janus II).  Ribosomal depletion via hybridization and cDNA synthesis are performed using the TruSeq Stranded Total RNA LP Globin kit as outlined by the manufacturer (Illumina, cat#20020612).  All steps are automated on the Perkin Elmer Sciclone NGSx Workstation to reduce batch to batch variability and to increase sample throughput.  Final RNASeq libraries are quantified using the Quant-it dsDNA High Sensitivity assay, and library insert size distribution is checked using a fragment analyzer (Advanced Analytical; kit ID DNF474).  Samples where adapter dimers constitute more than 4% of the electopherogram area are failed prior to sequencing.  Technical controls (K562,Thermo Fisher Scientific, cat# AM7832), are compared to expected results to ensure that batch to batch variability is minimized.  Successful libraries are normalized to 10nM for submission to sequencing.