RNA-seq of mus musculus:adult female small intestinal epithelial cells
RNA was extracted from FACS-sorted IECs using Trizol Reagent (Invitrogen, USA) by following the manufacturers procedure. Micro-RNA (mRNA) was isolated from total 1g RNA by using oligodT. After removal of rRNA, mRNAs were reverse-transcribed to generate single-stranded cDNA using random hexamer and reverse transcriptase, followed by double-stranded cDNA synthesis. Double-stranded cDNA was fragmented to the appropriate size and used in a standard Illumina library preparation involving end-repair, A-tailing and adapter ligation, and PCR amplification. After quantification of library using KAPAlibrary quantification kit, RNA-sequencing library was sequenced on a NovaSeq (Illumina, USA) followed by cluster generation. Purified total RNA and RNA-sequencing were performed by Theragen Etex (Korea).