GSM3132629: Con DG 3; Mus musculus; RNA-Seq
For samples 1-12, total RNAs were isolated using Direct-zol RNA Extraction Kit (Zymo Research). Library preparation and sequencing for RNA samples were performed at the Genomic Services Lab at HudsonAlpha Institute for Biotechnology (Huntsville, AL). Briefly, the quality of the total RNA was assessed using the Agilent 2100 Bioanalyzer. Two rounds of polyA+ selection was performed for RNA samples, followed by conversion to cDNAs. The mRNA library generation kits (Agilent, Santa Clara, CA) was used generate sequencing libraries per manufacturer's instructions. The indexed DNA libraries were quantitated using qPCR in a Roche LightCycler 480 with the Kapa Biosystems kit for library quantitation (Kapa Biosystems, Woburn, MA) prior to cluster generation. Clusters were generated to yield approximately 725 K to 825 K clusters/mm2. Cluster density and quality were determined during the run after the first base addition parameters were assessed.