title: GSM2028072: D2 Female3; Mus musculus; RNA-Seq
library_construction_protocol: The hippocampus (from both sides of the brain) was dissected and rapidly frozen in liquid nitrogen and stored at -80 degrees. Total RNA was extracted from the entire hippocampus from each animal using TRI Reagent (Sigma-Aldrich) according to manufacturer’s directions. RNA-Seq samples were prepared for each animal using the protocols of Mortazavi et al., 2008 and Chang et al., 2011.  The External RNA Controls Consortium (ERCC) RNA Mix 1 and Mix 2 Spike-in Controls (ERCC ExFold RNA Spike-In Mixes Ambion, catalog #4456739) were diluted 1:10 and spiked in to male and female libraries, respectively, just after mRNA isolation according to manufacturer’s suggestions. RNA-Seq sample quality was examined using Agilent High Sensitivity DNA Bioanalyzer Chips (Agilent Technologies 5067-4626) and quantified by KAPA Library Quantification Kits for Illumina sequencing platforms (KAPA Biosystems KK4824).Three libraries were generated for males and females from each strain. Barcodes were added to each library in order to combine multiple libraries in one sequencing lane. We multiplexed six libraries at a concentration of 12 pM and those were sequenced on an Illumina HiSeq 2500.