Circulating small RNAs in mouse obesity

Adult male C57BL/6 mice of similar age were kept for about 3 months on ad libitum high-fat or control diet (Bioserv products S3282 and F4031, with 60% and 10% fat calories respectively). Blood was collected by cardiac puncture into EDTA-containing tubes on ice. Plasma was isolated after centrifuging blood within 1 hour of collection and stored at -80 deg. C. Total RNA was isolated from plasma within a month of collection using Norgen Biotek's EXTRAClean Plasma/Serum RNA Purification Kit (product 73700) and sequencing libraries were prepared using Norgen Biotek's Small RNA Library Prep Kit for Illumina (product 63600). Libraries were stored at -80 deg. C for about 3 months before they were sequenced together on Illumina NextSeq 550 instrument using NextSeq 500/550 High Output Kit v2. All work was performed in one batch.