GSM7843365: Intestine, WT, fasted and refed, untreated, 2H [J20]; Mus musculus; RNA-Seq
RNA was extracted from whole tissue proximal intestine samples using the AllPrep DNA/RNA/Protein Mini Kit (80004, Qiagen) according to the manufacture's protocol. RNA pellet was resuspended in water, and concentrations were measured using a NanoDrop. Samples were submitted to Novogene and underwent sample quality control, library preparation, sequencing, and data quality control. For sample quality control, RNA was quantified, and purity was determined using NanoDrop. Sample integrity was determined using bioanalyzer by Agilent. All samples passed Novogene quality control. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis using dTTP for non-directional library. Next, the sample was assessed by Qubit and real-time PCR (RT-PCR) for quantification and bioanalyzer for size distribution detection. The clustering of index-coded samples was performed according to the manufacturer's instructions. After clustering generation, the library preparations were sequenced on Illumina platform, NovaSeq PE150 sequencing at 20 million reads per sample, and paired-end reads were generated.