Differential miRNA profile in the muscular and the visceral adipose tissue of morbidly obese patients undergoing bariatric surgery
Differential miRNA profile in the muscular and the visceral adipose tissue of morbidly obese patients undergoing bariatric surgery
Fresh visceral adipose tissue and skeletal muscle were obtained simultaneously during the bariatric surgery procedure under sterile conditions and immediately transported to the laboratory. A laminar flux cabin was used for  tissue manipulation to avoid contamination and blood vessels were cleaned out by using tweezers and scissors. Then, samples were frozen at -80oC immediately total RNA of each sample was extracted from 50-100 mg of tissue by using trizol reagent. Total RNA of each sample was used to prepare the miRNA sequencing library. After the completed libraries were quantified with Agilent 2100 Bioanalyzer, the DNA fragments in the libraries were denatured with 0.1M NaOH to generate single-stranded DNA molecules, captured on Illumina flow cells, amplified in situ and finally sequenced for 51 cycles on Illumina NextSeq 500 according to the manufacturer's instruction. The libraries were denatured as single-stranded DNA molecules, captured on Illumina flow cells, amplified in situ as clusters and finally sequenced for 51 cycles on Illumina NextSeq per the manufacturer's instructions.