Illumina NovaSeq 6000 paired end sequencing; MYDIASECRET: RNA-Seq of human skeletal muscle biopsies before and after bariatric surgery
MYDIASECRET: RNA-Seq of human skeletal muscle biopsies before and after bariatric surgery
Muscle biopsies taken under general anesthesia before surgery and under local anesthesia after surgery Frozen muscle samples were placed in Lysing matrix D tubes (MP Biomedical, France) containing 1 mL of Tripure™ Isolation Reagent (Roche Diagnostics, Germany) on ice and homogenized using the FastPrep-24™ Classic bead beating grinder and lysis system (MP Biomedical, France). Samples were homogenized 3 × 1 min, at 6 m/s, with 5-min rest on ice between rounds. Total RNA was isolated according to the manufacturer's instructions. High-throughput RNA-sequencing was performed on muscle biopsies. Total RNA was extracted from muscle samples as described previously. RNA integrity number (RIN ≥ 7) was checked using Bioanalyzer 2100 (Agilent, USA) and RNA 600 NanoChips before the preparation of libraries. Messenger RNAs were selected using KAPA mRNA HyperPrep Kit (Roche Sequencing Store, Roche, Switzerland) and reverse transcribed into complementary DNA.