title: 16S rRNA sequencing of gut microbiota from adult male Mus musculus intestinal content
design: Microbial DNA was extracted from mouse colon contents using the E.Z.N.A.? DNA Kit following the manufacturers protocol. DNA quality was assessed by 1% agarose gel electrophoresis to check integrity and quantified using a Qubit 4 Fluorometer. The V3-V4 hypervariable regions of the bacterial 16S ribosomal RNA gene were amplified by PCR with the following primers: forward primer 5-CCTAYGGGRBGCASCAG-3 and reverse primer 5-GGACTACNNGGGTATCTAAT-11. The PCR products were purified, quantified, and used for library construction.