title: Illumina HiSeq 4000 sequencing; RNA-seq of organoid derived monolayers (inflamed and non-inflamed) of UC patients and non-IBD controls, stimulated with microbiota of patients or a healthy donor
design: RNA-seq of organoid derived monolayers (inflamed and non-inflamed) of UC patients and non-IBD controls, stimulated with microbiota of patients or a healthy donor
library_construction_protocol: Confluent organoid derived monolayers of 8 UC patients and 8 non-IBD controls were stimulated for 6 hours with either control condition, UC microbiota or healthy donor microbiota, both with and without prior exposure to an inflammatory mix for 24 hours. Epithelial cells were frozen in lysis buffer at -80 after 6 hours co-culture with microbiota.   Total RNA was extracted using the Promega ReliaPrepâ„¢ miRNA Cell and Tissue Miniprep System. RNA was quantified by Nanodrop and the integrity was evaluated using Agilent Bio-analyzer 2100.   RNA libraries were prepared with the TruSeq Stranded mRNA library kit (Illumina, San Diego, USA).