GSM5989788: Sorted p16+ Stromal cells (CD45-CD31-CD29+)3; Mus musculus; RNA-Seq
After collection of tissue samples, samples were submerged in TRIzol reagent, and finely minced and grinded. For in vitro cultures, media was aspirated from tissue culture plates and TRIzol added according to the manufacturers recommended volume. The TRIzol was repeated pipetted across the dish. RNA isolation for both tissue and culture samples was then performed using Qiagen RNeasy Mini Kits following TRIzol extraction. After isolation of RNA as described previously, cDNA synthesis and library preparation were performed using the TruSeq RNA Library Prep Kit V2. The Agilent BioAnalyzer system was used to perform quality control on samples before and during library preparation. RNA samples with RIN scores below 7 post-cDNA synthesis were removed from analysis. Libraries were pooled and sequenced with paired-end sequencing to a depth of 50bp for each end at a targeted depth of 30M unique reads per sample.