GSM6926366: C57BL/6J_M_Saline_nucleus accumbens_1; Mus musculus; RNA-Seq
Flash frozen tissues were homogenized on ice in RLT buffer (Qiagen) containing 40 mM DTT with a Tissue-Tearor (BiosSpec Products, Inc) at low speed (setting 3) for 10 s. The resulting lysate was extracted with an equal volume of acid-phenol:chloroform (Thermo Fisher, cat# AM9722) and RNA was further purified from the aqueous phase with RNeasy mini kit (Qiagen) following the manufacture's recommendations. Samples were DNased (DNA-free DNA removal kit, Thermo Fisher) and RNA concentrations were determined using a Qubit Fluorimeter (Thermo Fisher). DNA-free total RNA was sent to Psomagen (Rockville, MD) for mRNA-seq. The 32 samples were sequenced to an average read count depth of 14.89 ± 2.9 million. Libraries were constructed using the Illumina TruSeq Stranded mRNA Library Prep Kit