GSM6310669: iBAT, cold, 3 days, rep 2; Mus musculus; RNA-Seq
Mice were euthanized by CO2 asphyxiation and cervical dislocation. The interscapular brown adipose tissue (iBAT) was surgically removed and placed into PBS on ice. iBAT was trimmed to remove interscapular white adipose tissue and flash frozen with liquid nitrogen. One iBAT pad was added to QIAzol reagent (QIAGEN) and RNA was isolated following the standard protocol for the Direct-zol RNA Miniprep kit (Zymo Research). RNA quality was assessed using the Agilent 2200 TapeStation (Agilent Technologies). Transcriptome profiles were generated using the QuantSeq 3'mRNA-Seq Library Prep Kit FWD for Illumina (Lexogen) with 250 ng of total RNA. Libraries were sequenced on an Illumina NovaSeq 6000 using a SP 100 cycle NovaSeq flow cell with read length configuration 76 bp with dual indexing: 12 bp for i7 and i5 indexing.