RNAseq of Caeca samples from conventional (CV) and germ-free (GF) chickens at 21 days of age
RNAseq of Caeca samples from conventional (CV) and germ-free (GF) chickens at 21 days of age
At day 21 post-hatching, CV and GF birds (n=8 per group) were sacrificed by cervical dislocation followed by blood withdrawal from the occipital venous plexus. Blood samples were stored in heparin-coated tubes for subsequent plasma preparation. Caeca samples were immediately collected from each chicken, snap frozen in liquid nitrogen and stored at -80°C until further use for RNA extraction. Total RNA was extracted using the NucleoSpin RNA II kit (Macherey-Nagel, Germany) with rDNAse treatment to eliminate contaminant DNA. RNA quality was checked by the Bioanalyzer 2100 system using the RNA Nano 6000 Assay kit (Agilent Technologies, CA, USA).  The NEBNext® Ultra™ II RNA Library Prep Kit for Illumina® was used to prepare the library. RNA with PolyA selection.