Microbiota profiling of caecal contents from conventional chickens beetween 7 and 21 days of age
Microbiota profiling of caecal contents from conventional chickens beetween 7 and 21 days of age
At days 7, 14 and 21 post-hatching conventional (CV) chickens were sacrificed by cervical dislocation followed by blood withdrawal from the occipital venous plexus. Caecal contents were immediately collected from each chicken, snap frozen in liquid nitrogen and stored at -80°C until further use for DNA extraction and 16S sequencing.   Total DNA was extracted from caecal contents using the Dneasy PowerSoil kit (Qiagen, UK) following the manufacturer's protocol. 16S (V3-V4 region) amplification was performed as described by Zheng et al (An accurate and efficient experimental approach for characterization of the complex oral microbiota. Microbiome 3, 48, 2015). In the limited cycle 2nd round PCR, sample-specific barcodes were added for multiplexing.