GSM5060716: OBM4_S19; Homo sapiens; RNA-Seq
Approximately 500 mg of frozen SAT was homogenized in RLT buffer (Qiagen). RNA from SAT was extracted using the RNeasy Mini Kit (Qiagen) according to the manufacturer protocol and samples were then treated with the RNase-Free DNase Set (Qiagen). Concentration and quality of the extracted RNA were determined by the NanoDropH ND-1000 spectrophotometer (NanoDrop Technologies, USA) and RNA integrity verified by gel-electrophoresis. RNA-seq libraries were prepared with the CORALL Total RNA-Seq Library Prep Kit (Lexogen) using 150 ng total RNAs from 5 healthy women, 5 obese women, 5 obese women with T2D and 5 obese men. The RiboCop rRNA Depletion Kit (Lexogen) was used to remove rRNA. Qualities of sequencing libraries were assessed with D1000 ScreenTape Assay using the 4200 TapeStation System (Agilent) and quantified with Qubit™ dsDNA HS Assay Kit (Invitrogen). RNA processing was carried out using Illumina NextSeq 500 Sequencing.