GSM6329631: WT non-SVE 2; Mus musculus; RNA-Seq
Once entrained, animals in the SVE cohort were culled 2h after the time at which the wheel became available to exercise in (ZT14). For animals in the nSVE cohort, they were culled at one timepoint (CT14). The brain was removed, flash frozen on dry ice, and stored at -80 C. Brain sections of 20µm thickness were cut using a cryostat (Leica CM3050 S) onto PEN-membrane slides (Leica Biosystems, Germany) and then stored at -80 °C. Slides were then defrosted and to aid in identifying the SCN and its anatomical boundaries, sections were stained with 1% cresyl violet (Merck) in 70% ethanol. The SCN was identified and extracted on a laser-capture microscope (LCM) system (Leica DM6000 B) and stored in lysis buffer (Promega) prior to RNA extraction. RNA extraction was carried out with ReliaPrep Kit (Promage, UK). Total RNA was checked using RNA 6000 Pico Kit (Agilent Technologies, USA) and amplified with Sense Total RNA-Seq Library Prep Kit (Lexogen, USA). RNA sequencing was done by paired-end sequencing on an Illumina HiSeq 4000 NGS platform (Genomic Technologies Core Facility, University of Manchester, UK)