GSM8758572: Control,D30,rep2; Mus musculus; RNA-Seq
The jejum tissues of the mice were dissected according to their anatomic features and stored immediately in RNAlater solution (QIAGEN) in RNAse-free 1.7-ml tubes (Denville Scientific, Holliston, MA) at -80°C to stabilize and protect the integrity of RNA. For each sample, total RNA was extracted from ~1 cm of ileum. Prior to the extraction, 500 µl of ice-cold RNase free PBS was added to the sample. To remove the RNAlater, the mixture was centrifuged at 8,000xg for 10 min and the resulting pellet resuspended in 500 µl ice-cold RNase-free PBS with 10 µl of β-mercaptoethanol. Ribosomal RNA depletion and library construction were performed using the RiboZero Plus kit and TruSeq stranded mRNA library preparation kit Ribosomal RNA depletion and library construction were performed using the RiboZero Plus kit and TruSeq stranded mRNA library preparation kit (Illumina) according to the manufacturer's recommendations and sequenced using Illumina NovaSeq 6000 S2 flow cell