16S rRNA sequencing of gut microbiota from adult male Mus musculus intestinal content
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-7. The PCR products were purified, quantified, and used for library construction.