GSM9238084: Normal mouse, High-fat diet (HFD) + water, 10 weeks, rep4; Mus musculus; RNA-Seq
Total RNA was extracted from the tissue using TRIzol®Reagent according the manufacturer's  instructions. Then RNA quality was determined by 5300 Bioanalyser (Agilent) and quantified using the ND-2000 (NanoDrop Technologies). Only high-quality RNA sample (OD260/280=1.8~2.2, OD260/230≥2.0, RQN≥6.5, 28S:18S≥1.0, >1μg) was used to construct sequencing library. RNA purification, reverse transcription, library construction and sequencing were performed at Shanghai Majorbio Bio-pharm Biotechnology Co.,Ltd. (Shanghai,China) according to the manufacturer's instructions. The XXRNA-seq transcriptome librariy was prepared following Illumina®Stranded mRNAPrep, Ligation (SanDiego, CA) using 1μg of total RNA. Shortly, messenger RNA was isolated according to polyA selection method by oligo(dT) beads and then fragmented by fragmentation buffer firstly. Secondly double-stranded cDNAwas synthesized using a SuperScript double-stranded cDNAsynthesis kit (Invitrogen, CA)with random hexamer primers.Then the synthesized cDNA was subjected to end-repair, phosphory lation and adapter addition according to library construction protocol. Libraries were sizeselected for cDNA target fragments of 300bp on 2% Low Range Ultra Agarose followed by PCR amplified using Phusion DNA polymerase (NEB) for 15 PCR cycles. After quantified by Qubit4.0, these quencing library was performed on NovaSeq X Plus platform (PE150) using NovaSeq Reagent Kit.