title: GSM8548621: Quadriceps_CONVR_6845; Mus musculus; RNA-Seq
library_name: GSM8548621
library_construction_protocol: At ~12 weeks of age, all mice were sacrificed for the extraction of brain, colon, liver, gonadal white adipose tissue (gWAT), and quadriceps tissues. RNA was isolated from brain, colon, liver, and quadriceps via the “RNeasy mini kit” (Qiagen, Hilden, Germany) according to the manufacturer's protocol, while RNA from gWAT was isolated using the TRIZOL method as previously described 98. Briefly, 1 ml TRIzol was added to 50–75 mg pestle-homogenized tissue followed by vortexing, a 5-min incubation at room temperature, and addition of 200 μl chloroform. After mixing, further incubation at room temperature for 2–3 min and centrifugation (12.000 g) at 4 °C for 5 min, the clear supernatant was mixed with 500 μl isopropanol followed by incubation at room temperature for 10 min. After further centrifugation (12.000 g) at 4 °C for 10 min, the supernatant was discarded and the pellet washed with 1 ml cold 75% EtOH followed by vortexing and centrifugation (7.500 g, 4 °C, 5 min). The pellet was dried and dissolved in RNase-free water. All animal protocols were approved by the Gothenburg Animal Ethics Committee (vote #2652-19). RNA was reverse-transcribed into Illumina shotgun sequencing libraries with TruSeq RNA stranded kit and polyA enrichment following the manufacturer's protocol (Illumina, San Diego, CA) at the Competence Centre for Genomic Analysis (Kiel, Germany) and sequenced for 2×100 cycles in paired-end mode on an Illumina NovaSeq6000 machine.