title: GSM8120904: aorta, HFD, rep1; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was extracted using Trizol reagent kit (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's protocol. RNA quality was assessed on an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA) and checked using RNase free agarose gel electrophoresis. After total RNA was extracted, eukaryotic mRNA was enriched by Oligo(dT) beads. Then the enriched mRNA was fragmented into short fragments using fragmentation buffer and reversly transcribed into cDNA by using NEBNext Ultra RNA Library Prep Kit for Illumina (NEB #7530,New England Biolabs, Ipswich, MA, USA).The purified double-stranded cDNA fragments were end repaired, A base added, and ligated to Illumina sequencing adapters. The ligation reaction was purified with the AMPure XP Beads (1.0X). Ligated fragments were subjected to size selection by agarose gel electrophoresis and polymerase chain reaction (PCR) amplified. NEBNext Ultra RNA Library Prep Kit RNA-seq Principal component analysis (PCA) was performed with R package gmodels in this experiment. RNAs differential expression analysis were performed by DESeq2 software between two groups. The genes/transcripts with parameter of false discovery rate (FDR) below 0.05 and absolute fold change≥2 were considered differentially expressed gene/transcripts.