GSM8729968: BU-LF-65; Mus musculus; RNA-Seq
Total RNA was extracted from the cortex using Trizol reagent and cleaned using RNeasy spin columns with DNase treatment to remove contaminating genomic DNA. RNA libraries were prepared from 200 ng of total RNA using the TruSeq Stranded RNA Sample Prep Kit (Illumina, San Diego, CA) according to the manufacturer's instructions, employing poly-A mRNA enrichment using oligo-dT magnetic beads. The final adaptor-modified cDNA fragments were enriched by 15 cycles of PCR using Illumina TruSeq PCR primers. The concentration and size distribution of the completed libraries were determined using an Agilent Tape Station, (Agilent, Santa Clara, CA) and Qubit fluorometer (Invitrogen, Carlsbad, CA). Libraries from all samples were sequenced on Illumina's NovaSeq 6000 at 50 million fragment reads/sample following Illumina's standard protocol on two S2 flow cells. S2 flow cells were sequenced at 101 x 2 paired end reads using a NovaSeq S2 sequencing kit, NovaSeq Control Software v1.7.0 and base-calling was analyzed using Illumina's RTA version 3.4.4.