title: GSM8100709: sham,biol rep 3,bulk_RNA_seq; Mus musculus; RNA-Seq
library_construction_protocol: Mice were treated with sham, TPN, or starvation for 24 h, and then 5 cm of jejunal tissue was harvested and longitudinally opened. After the RNA samples were tested, eukaryotic mRNA was enriched with magnetic beads with Oligo(dT) . Subsequently, mRNA was interrupted into short fragments by adding fragmentation buffer. One-stranded cDNA was synthesized by using six-base random primers (random hexamers) with mRNA as template, and then two-stranded cDNA was synthesized by adding buffer, dNTPs, and DNA polymerase I and RNase H. The purified double-stranded cDNA was first subjected to end repair, addition of A-tail and ligation of sequencing junctions, followed by fragment size selection with AMPure XP beads. Finally, PCR amplification was performed and the PCR products were purified by AMPure XP beads to obtain the final library. After the library was constructed, Qubit 2.0 was used to perform preliminary quantification and dilution of the library, followed by Agilent 2100 to detect the insert size of the library, and after the inserts met the expectation, Q-PCR was used to accurately quantify the effective concentration of the library to ensure the quality of the library.