GSM4563881: ShamP1_Podo_1; Mus musculus; RNA-Seq
RNA was isolated with trizol according to manufacturer's instructions with glycogen included as a carrier. Each sample was treated with DNase I (Roche) for 10 minutes and then RNA was further column purified with MinElute RNeasy kit (#28004, Qiagen). RNA quality was ascertained by RNA-Pico on Agilent Bioanalyzer. RNA-seq libraries were constructed using Ovation RNA-Seq System V2 followed by Ovation Ultralow System V2 using Nugen protocols M01114 v5 and M01379v3.1. Libraries were quantitated by Qubit and size profile determined using Bioanalyzer. All libraries had a mean size of ~300 bp and were similar concentrations (~10 ng/ul) Libraries were finally quantitated by qPCR and 2 pools prepared in equimolar ratios for sequencing. 200 pM of each pool based upon qPCR was used for clustering. Libraries were sequenced at 80 bp single base pair reads using Illumina HiSeq 3000.