snRNA-seq of mus musculus: adult female nucleus accumbens
Briefly, 3 mice with or without VLTE were euthanized and perfused intracardially with PBS; and brains were then collected and snap frozen with liquid nitrogen. The nucleus accumbens was micro-dissected and approximately 10 g of tissue was used for single nuclei isolated as previously described (Schmutz, 2021) as well as nuclei fixation using the Evercode Nuclei Fixation v3 kit (Parse Biosciences). The number of nuclei isolated for each sample was then counted by trypan blue using a hemocytometer and a total of 2,500 nuclei from 3 samples per group (+/- VLTE) were used to construct the snRNAseq library using the Whole Transcriptome Mini v2 kit (Parse Biosciences) in accordance with the manufacturers instructions. One of the two sub-libraries generated was then sequenced using NextSeq 2000 with P3 XLEAP (100 cycles) targeting approximately 30,000 reads per nuclei.