RNA-Seq of mus musculus: adult male hippocampus
Fresh tissues of hippocampus of LC+Veh, HF+Veh and HF+DI mice were collected to analyze the whole profile of transcriptome. Total RNA was extracted using Trizol reagent kit (Invitrogen, Carlsbad, CA, USA) according to the manufacturers protocol and mRNA was enriched by Oligo(dT) beads. Then the enriched mRNA was fragmented into short fragments using fragmentation buffer and reversly transcribed into cDNA. 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.The resulting cDNA library was sequenced using Illumina Novaseq6003 by Gene Denovo Biotechnology Co. (Guangzhou, China).