Composition and diversity of the bacterial communities. DNA in
each ileal and colonic chyme sample was extracted according to the
manufacturers protocol (Bocai Biology). The V3 and V4 regions of the
16S ribosomal DNA gene were chosen for PCR. The primers were
338F (5#ACTCCTACGGGAGGCAGCA-3#) and 806R (5#GGACTACHVGGGTWTCTAAT-
3#). The procedure to obtain amplicons
was previously described, with modifications The products were
examined on a 2% (wt:vol) agarose gel. The amplicons from 6 replicates/
treatment were blended in equimolar ratios on the basis of concentration.
The blended samples were sent out for MiSeq Next-Generation Sequencing
System on an Illumina MiSeq PE300 platform at the Majorbio Bio-
Pharm Technology Company. Only sequences >50 bp were used for
phylotype analysis at the 0.03 operational taxonomic unit level.