title: GSM1470818: DARPA_46_CCGTCC_L003_R1; Mus musculus; RNA-Seq
library_construction_protocol: RNA samples were extracted using TRIzol® reagent (Life Technologies, Grand Island, NY) from the striatum of a random subset of 100 F2 mice, and were then shipped to the Genomics Core Facility at the Icahn School of Medicine at Mount Sinai. Sample integrity was assessed using either an Agilent 2100 Bioanalyzer with the RNA 6000 Nano assay or an Agilent 2200 TapeStation with the R6K ScreenTape (Agilent, Santa Clara, CA). The average RIN values were above 9 for all processed total RNA samples. The standard TruSeq RNA Sample Prep Kit v2 protocol (Illumina, San Diego, CA) was used to prepare the sequencing library. Briefly, the cDNA was synthesized from poly-A-selected and then fragmented total RNA using random hexamers, followed by end-repair and ligation with appropriate sequencing adaptors. The libraries were then subject to size selection and was purified using AMPure XP beads (Beckman Coulter, Brea, CA). Barcode bases (6 bp) were introduced at one end of the adaptors during PCR amplification steps. The size and concentration of the cDNA libraries was measured by Bioanalyzer and Qubit fluorometry (Life Technologies, Grand Island, NY) before sequencing. The cDNA libraries were sequenced on the Illumina HiSeq 2500 System with 100 nucleotide single-end reads, according to the standard manufacturer's protocol (Illumina, San Diego, CA).