RNA-Seq of the posterior subcutaneous fat of mice after 3-week treadmill training versus sedentary control
RNA-Seq of the posterior subcutaneous fat of mice after 3-week treadmill training versus sedentary control
Tissue samples were removed, weighed, snapped frozen in liquid nitrogen, and stored at -70°C. Total RNA was isolated using TRI reagent (Sigma-Aldrich, Taufkirchen, Germany). DNA traces were removed by DNaseI-treatment (Baseline-Zero DNase; Biozym, Hessisch Oldendorf, Germany), and the RNA was cleaned-up using the RNA Clean&Concentrator-25 Kit (Zymo Research, Freiburg, Germany). RNA quantity was determined using a Qubit fluorometer (ThermoFisher Scientific, Germany), and integrity was checked on an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). 1 µg of total RNA with RIN > 8 was used for library preparation using a TruSeq Stranded mRNA Kit with 11 cycles of PCR-amplification according to the manufacture's recommendation (Illumina).