GSM5289470: WT-sedentary group mouse 3; Mus musculus; RNA-Seq
Hippocampi were micro-dissected, flash frozen on dry ice, and the nuclei were isolated. The isolated nuclei were stained woth Vybrant Ruby dye and Ruby+/GFP+ nuclei were FACS-sorted. The cDNA was synthesized from sorted cells using Takara SmartSeq v4 reagents according to manufacturer's protocol with modification: 1μl of 2X Reaction buffer was used with 1μl of CDS primer and 18 cycles of PCR were used for the cDNA amplification step. Full length cDNA was fragmented to a mean size of 150bp with a Covaris M220 ultrasonicator and Illumina libraries were prepared from 2ng of sheared cDNA using Rubicon Genomics Thruplex DNAseq reagents according to manufacturer's protocol. Uniquely indexed libraries were pooled in equimolar ratios and sequenced with single-end 75bp reads on an Illumina NextSeq500 run by the Dana-Farber Cancer Institute Molecular Biology Core Facilities.