GSM8765965: Cortex.B4B9; Mus musculus; RNA-Seq
Mice were anesthetized via inhalation of isoflurane and euthanized by cervical dislocation. Blood was collected via cardiac puncture followed by transcardial perfusion of 10 mL PBS to wash tissues. Tissues were removed and washed in PBS twice before being flash frozen in liquid nitrogen. Following collection, tissues were stored at -80ºC before further processing. For RNA extraction, RNeasy Mini kits were used (QIAGEN #74104). After extraction, RNA was stored at -80ºC until further processing. Libraries were prepared using the NEBNext UltraTM II kit. In brief 500 ng of RNA per sample un-derwent Poly(A) selection before fragmentation and random priming followed by first and second strand cDNA synthesis. Following cDNA synthesis, NEBNext Multiplex Oligos for Illumina (E6440S) were ligated before PCR enrichment. All libraries were pooled to a final concentration of 1nM and sequenced on a NovaSeq 6000.