title: GSM9470406: Hind 3month KO Mouse 99_3; Mus musculus; RNA-Seq
library_construction_protocol: Each brain region from each animal was then separately prepared for bulk RNA sequencing. RNA was isolated from tissues using the Qiagen RNeasy Mini Kit per manufacturer's instructions. . Bulk sequencing libraries were prepared using Illumina TruSeq Stranded mRNA Kit (Catalog #20020595) per manufacturer's instructions. 300 ng of total RNA from each sample was used and the concentration of resulting cDNA library was measured with High Sensitivity DNA chips (Agilent) using a 2100 Bioanalyzer Instrument (Agilent). A 5 nM normalized library was pooled, and sequencing was performed on a NovaSeq X 25B (Illumina) with 50 bases each for reads 1 and 2 and 8 bases each for index reads 1 and 2. The depth of sequencing for each individual library ranged from 30.5 million to 645 million before downsampling, with a median of 49 million reads.