title: RNA-seq of human with nonalcoholic fatty liver disease against untreated controls
design: RNA-seq of human with nonalcoholic fatty liver disease against untreated controls
library_name: Sample 23_p
library_construction_protocol: Patients and tissue collection. for HC, presence of a normal liver (no steatosis or cirrhosis) on ultrasound evidence; for NAFLD was based on ultrasound evidence of fatty liver in the absence of other (viral, alcohol, metabolic) causes. Liver tissue was collected by a wedge liver biopsy from free edge of the liver lobe during laparoscopic cholecystectomy. The study protocol received institutional review board (IRB) approval before the beginning of the study, in accordance with The Code of Ethics of the Declaration of Helsinki, and informed consent was obtained from all patients. tissue samples were immediately stored in RNA-later for 24 hours at 4℃, and subsequently at -80℃. RNA isolation and quality controls. Approximately 10-20 mg of liver tissues were homogenized in 1 mL Trizol (Thermo Fisher Scientific, shanghai, China) on ice, incubated for 5 min, then added 200 μL chloroform. After low-temperature centrifugation, 500 μL isopropanol was used to precipitate RNA. The precipitate was washed with 75% ethanol (DEPC water configuration), and then dissolved in 30 μL DEPC water (shanghai Generay Biotech Co., Ltd, shanghai, China) after evaporating all ethanol. The RNA concentration, purity and integrity were measured by Agilent 2100 Bioanalyzer (Agilent Technologies Inc., Beijing, China). RNA concentration, purity and RNA integrity number (RIN) were measured using an Agilent 2100 Bioanalyzer before sequencing. 2 μg of RNA meeting the requirements of RIN ≥ 7 and 28S/18S ≥ 1.0 were taken for library construction.