alias: GSM2319214
title: GSM2319214: stress-HT rep1 RNA-seq; Mus musculus; RNA-Seq
library_construction_protocol: Mice were sacrificed (2 hours after lights on) and whole brains were extracted and immediately flash frozen in 2-methylbutane and dry ice. Striatum tissue was excised by micropunch (1.53 to -0.95 mm posterior to bregma) and approximately 30 milligrams of tissue was homogenized with glass beads (Sigma) and DNA/RNA were extracted using AllPrep DNA/RNA mini kit (Qiagen). 5hmC-enriched libraries were generated using the NEBNext ChIP-Seq Library Prep Reagent Set for Illumina sequencing, according to the manufacturer’s protocol. Briefly, the 5hmC-enriched DNA fragments were purified after the adapter ligation step using AMPure XP beads (Agencourt A63880). Approximately 100 ng of total RNA was used for sequence library construction following instructions of NuGen mRNA sample prep kit (cat# 0348). In brief, total RNA was copied into first strand cDNA using reverse transcriptase and random primers. This was followed by second strand cDNA synthesis using DNA Polymerase I and RNaseH. These cDNA fragments went through an end repair process, the addition of a single ‘A’ base, and then ligation of the adapters. These products were gel purified and enriched with PCR to create the final cDNA libraries.
identifiers.namespace: GEO
identifiers.id: GSM2319214