Name: GSM7054405
Instrument: Illumina NovaSeq 6000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC SINGLE CELL
Selection: cDNA
Layout: PAIRED
Construction protocol: Paired samples of omental and subcutaneous WAT were obtained from female patients with obesity undergoing different bariatric surgery procedures (Roux-en-Y Gastric Bypass and sleeve gastrectomy). After collection, samples were washed using PBS, placed on ice until the end of the surgery procedure, snap-frozen in liquid nitrogen and stored at -80°C for later use. 50 mg of fresh or frozen WAT was first minced into 1-3 mm pieces and then homogenized on ice in 0.1% CHAPS in CST buffer supplemented with 0.2U/μl RNAase inhibitor (RI) using a Dounce homogenizer. After homogenization, samples were left on ice for five minutes following which PBS supplemented with BSA and 0.2U/μl RI was added to obtain a final concentration of 1% BSA. The lysates were filtered through 40 μm cell strainers and centrifuged at 500 x g for five minutes at 4 °C. The nuclei pellets were resuspended with 1% BSA in PBS supplemented with 0.2U/μl RI and centrifuged again at 500 x g for five minutes at 4 °C. This step was repeated once more. After the final resuspension, nuclei were filtered through 20 μm cell strainers and loaded directly on a 10x Chip G. 10X-libraries were prepared with the Chromium Single Cell v3.1 reagent kit following the manufacturer's protocol (10X Genomics). Suspensions containing around 1200 nuclei per μl were loaded on Chip G followed by reverse transcription to obtain cDNA, which subsequently was amplified and used for library construction. After preparation, the libraries were sequenced on a NovaSeq 6000 platform (Illumina). snRNA-seq