Illumina HiSeq 4000 sequencing; Identifying molecular features contributing to clinical heterogeneity in Crohn's disease using multi-omic data integration
Identifying molecular features contributing to clinical heterogeneity in Crohn's disease using multi-omic data integration
We isolated PBMCs and sorted monocytes and CD4 T cells through FACS prior to the initiation of biological therapy. All recruited patients had endoscopy proven disease at time of sample collection. Total RNA lysed, sorted CD4+ and CD14+ cells was extracted using the AllPrep DNA/RNA Mini kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions. The integrity and quantity of all RNA was assessed with a 2100 Bioanalyzer (Agilent, Waldbronn, Germany) and a Nanodrop ND-1000 spectrophotometer (Thermo Scientific, Waltham, Massachusetts, USA).  Library preparation was performed using the TruSeq Stranded mRNA protocol (Illumina, San Diego, USA) according to the manufacturer's instructions.