title: RNA-seq of ChI-D2RKO mouse nucleus accumbens following a progressive ratio schedule of reinforcement of intravenous cocaine or saline
design: RNA-seq of ChI-D2RKO mouse nucleus accumbens following a progressive ratio schedule of reinforcement of intravenous cocaine or saline
library_construction_protocol: Brains were rapidly dissected 1 hour after the end of the progressive ratio session and frozen in 2-methylbutane on dry ice.  Bilateral tissue punches of the ventral striatum were obtained and rapidly homogenized in TRIzol (Invitrogen; Cat# 15596026). Total RNA was extracted following the manufacturer's recommendation. Total RNA was monitored for quality control using the Nanodrop absorbance ratios for 260/280nm and 260/230nm. Total RNA was monitored for quality control using the Agilent Bioanalyzer Nano RNA chip and Nanodrop absorbance ratios for 260/280nm and 260/230nm. Library construction was performed according to the Illumina TruSeq® Stranded mRNA Sample Preparation Guide. The input quantity for total RNA was 500ng and mRNA was enriched using oligo dT magnetic beads. The enriched mRNA was chemically fragmented for three minutes. First strand synthesis used random primers and reverse transcriptase to make cDNA. After second strand synthesis the ds cDNA was cleaned using AMPure XP beads and the cDNA was end repaired and then the 3' ends were adenylated. Illumina barcoded adapters were ligated on the ends and the adapter ligated fragments were enriched by nine cycles of PCR. The resulting libraries were validated by qPCR and sized by Agilent Bioanalyzer DNA high sensitivity chip.