title: Illumina HiSeq 2500 paired end sequencing; RNA-seq bulk of CCR6+Th-cell subsets in Crohn's Disease, peripheral blood and intestinal lamina propria
design: RNA-seq bulk of CCR6+Th-cell subsets in Crohn's Disease, peripheral blood and intestinal lamina propria
library_construction_protocol: Peripheral blood mononuclear cells were magnetically enriched for CD4+ T-cells with a kit (Miltenyi). T-cells were stained with antibodies to CD4, IL-7R, CD25, CCR6, CXCR3 and CCR5 and purified on a FACS Aria (BD). CD4+IL-7R+CD25-/lo helper T-cells were purified as cTh17- (CCR6+CXCR3-CCR5-), pTh17- (CCR6+CXCR3-CCR5+), CCR5-Th1/17- (CCR6+CXCR3+CCR5-) or CCR5+Th1/17-cells (CCR6+CXCR3+CCR5+). RNA was isolated using mirVana Isolation Kit (ThermoFisher) Library construction was performed with the TruSeq RNA Sample Preparation Kit v2 (Illumina)