GSM7231223: NAc LSD 2 days replicate 1; Mus musculus; RNA-Seq
At P98-P112 mice were sacrificed, brains were rapidly removed and ~1mm thick coronal slice (n = 3 mice for each conditions) containing the nucleus accumbens were sectioned using a mouse brain matrix. To microdissect the NAc, slices were placed in a petri dish containing ice-cold, ACSF (125 mM NaCl, 2.5 mM KCl, 2 mM CaCl2, 1 mM MgCl2, 1.25 mM NaH2PO4, 10 mM glucose and 26 mM NaHCO3) supplement with RNase Inhibitor and oxygenated with carbogen gas (95% O2 and 5% CO2) to pH 7.3 - 7.4. The NAc was identified using the anterior commissure and other structural markers. Between each dissection, blades were replaced and all the instruments and the matrix were cleaned with a solution containing RNase inhibitor. Following dissection, tissue was immediately placed into 0.5 ml trizol and subjected to a 15 second burst with a tissue homogenizer to lyse the cells.  Samples were kept on ice prior to storage at -20C. Total RNA were extracted using the RNeasy Kit from Qiagen. The quality of purified RNA was assessed via both a nanodrop and 2100 Bioanalyzer from Agilent. Library preparation was performed using a TruSeq Stranded mRNA kit (Illumina) using the recommended protocol. Individual dual-indexed libraries were quality controlled, pooled, and sequenced on the NovaSeq 6000 platform on a single S1 flowcell to an average depth of 76,841,745 (± 8,066,939.82) paired-end 100bp reads per sample.