title: miRNA-seq of uninflamed colon region from TNBS treated rat
design: miRNA-Seq was performed by TruSeq Small RNA Library Preparation Kit (Illumina), according to the manufacturers instructions and sequenced on Illumina MiSeq instru-ment. Briefly, during 3 adapter ligation reactions achieved by the T4 RNA Ligase 2 (Dele-tion Mutant) enzyme, 1 g total RNA was used from each rat colon sample. After 5 adapter ligation by T4 RNA Ligase, reverse transcription was performed by SuperScript II Reverse Transcriptase (Thermo Fisher), and the constructs were amplified by PCR. Ampli-fication products were verified on TapeStation High Sense DNA tape (Agilent). cDNA contracts were purified and size selected by gel electrophoresis on 6 % Novex TBE gels (Thermo Fisher). To elute the DNA, gel slices were broken and incubated in elution buffer overnight. Gel slurry was removed by Ambion Spin Column, and the DNA was precipi-tated with isopropanol. Resuspended DNA libraries were quality checked on TapeStation High Sense DNA tape. Final libraries were quantified by Qubit (Thermo Fisher) and se-quenced on Illumina MiSeq instrument using the 50-base sequencing chemistry.