title: 16S rRNA V4 gene amplicon sequencing of DNA extraction negative control 3 (515F/806R) [JMF-2112-06-0053A]
design: Amplicon sequencing of DNA extraction negative control 3. Timed matings were set up in the same animal room to ensure that all mice were of the same age. The offspring were genotyped and weaned at 3 weeks of age. At weaning, the offspring from different litters were mixed and 3 CDHR5-proficient and 3 CDHR5-deficient mice from different litters were caged together until they were 8 weeks old. Fecal pellets were collected from 8 week old adult mice. After pellet collection, adult females were remixed and 3 CDHR5-proficient and 3 CDHR5-deficient mice were housed together per cage. Adult male mice could not be mixed but were split to have 3 mice per cage (either 2 CDHR5-proficient and 1 CDHR5-deficient mice or 2 CDHR5-deficient and 1 CDHR5-proficient mice) to minimize cage effects. Mice were treated with DSS in sterile autoclaved drinking water (female mice 2.0%, male mice 1.5% DSS) for 7 days, followed by a 7-day recovery period without DSS. Control mice received sterile autoclaved drinking water throughout the experiment. The weight and condition of the mice were monitored daily to ensure colitis. A second collection of fecal pellets was performed under a hood on day 14 after the recovery period and the pellets were immediately frozen in a pre-weighed Eppendorf tube on dry ice. Sequencing was performed as described in https://doi.org/10.3389/fmicb.2021.669776. {primers: GTGYCAGCMGCCGCGGTAA + GGACTACNVGGGTWTCTAAT, nucleic acid: DNA, flow cell identifier: K7H6J, barcoding: UDB-H12, plate-position: 1 A8, barcodes: ATATACCTGCGG + ATCACATTCTCC}.