title: GSM9684674: WT_rep2; Mus musculus; RNA-Seq
library_construction_protocol: Mice were deeply anesthetized with an inhalation anesthetic (isoflurane) prior to euthanasia by cervical dislocation. Total RNA was extracted from the whole left brain of 10-week-old mice using the RNeasy Plus Mini Kit (QIAGEN) according to the manufacturer's instructions. RNA integrity was evaluated using the Agilent Fragment Analyzer. RNA sequencing libraries were constructed using the NEBNext® Ultra™ II Directional RNA Library Prep Kit (New England Biolabs) according to the manufacturer's instructions. Polyadenylated mRNA was enriched from 400 ng of total RNA using oligo-dT magnetic beads, fragmented, and reverse-transcribed into first-strand cDNA using random hexamer primers. Following second-strand cDNA synthesis, end repair, A-tailing, adapter ligation, and size selection, the libraries were amplified and purified. Libraries were circularized using the MGIEasy Universal Library Conversion Kit (MGI Tech).