title: Illumina HiSeq 2500 sequencing; Transcriptional profiling of murine small intestinal epithelial cells stimulated with interleukin-28A
design: Transcriptional profiling of murine small intestinal epithelial cells stimulated with interleukin-28A
library_construction_protocol: Murine small intestinal organoids were stimulated with 1000ng/ml IL-28A or left untreated for 24h. Total RNA was isolated and subjected to RNA-Seq (4 stimulated vs 4 unstimulated). The quality of the isolated RNA was determined with a Qubit (2.0) Fluorometer (Life Technologies, California, USA) and a Bioanalyzer 2100 (Agilent, Waldbronn, Germany). The TruSeq RNA Sample Prep Kit v2 (Illumina, Inc, California, USA) was used. Briefly, total RNA samples (100-1000 ng) were poly A enriched and then reverse-transcribed into double-stranded cDNA. The cDNA samples was fragmented, end-repaired and polyadenylated before ligation of TruSeq adapters containing the index for multiplexing Fragments containing TruSeq adapters on both ends were selectively enriched with PCR. The quality and quantity of the enriched libraries were validated using a Qubit (2.0) Fluorometer (Life Technologies, California, USA) and a Bioanalyzer 2100.