title: GSM4816984: sample_12; Mus musculus; RNA-Seq
library_construction_protocol: Brain microdissections were collected from mice that were euthanized by cervical dislocation. Brains were coronally sectioned using a brain matrix into 1 mm-thick slices. A 14G blunt needle was used for microdissection of the vmPFC. Viral expression and placement were confirmed using an epifluorescence microscope. Total RNA was purified from the tissue using the RNeasy micro kit (Qiagen, Germantown, MD) and TriZol. Samples were treated with DNase I and concentration and purity were tested using NanoDrop (Thermo Fisher Scientific, Waltham, MA). Libraries were prepared from 7 individual mice per group using the TruSeq Stranded Total RNA Library Prep Gold (Illumina, San Diego, CA) from virally infected mPFC tissue from female mice at baseline or after 3 days of sCVS. Briefly, 1 µg ribosomal depleted RNA was converted to cDNA using random hexamers. Adapters were ligated and samples were size selected with AMPure XP beads (Beckman Coulter, Brea, CA). Barcode bases of 6 bp were introduced at one end of the adapters during PCR amplification steps. Quality and concentration of libraries were analyzed with a Bioanalyzer DNA 1000 chip (Agilent, Santa Clara, CA). 7 samples were pulled per lane for multiplexing and sequenced on an Illumina Hi-Seq machine with 260 bp paired-ends reads by Genewiz (South Plainfield, NJ) and gave rise to ~35 million reads per sample.