title: Bacterial 16S amplicon sequencing, mouse fecal sample Cx27_20, WT: CX3CR1DTR x CD11c cre -/- diphtheria toxin treated
design: Bacterial 16S regions were PCR amplified using primers 16S-27F AGAGTTTGATCMTGGCTCAG and 16S-BSR357R CTGCTGCCTYCCGTA. 16S amplicons were generated with 25 cycles using Invitrogen AccuPrime PCR reagents (Carlsbad, CA). Amplicons were then used in the second PCR reaction, using Illumina Nextera XT v2 (San Diego, CA) barcoded primers to uniquely index each sample and 2x300 paired end sequencing was performed on the Illumina MiSeq (Illumina, CA). DNA amplification PCR: Initial denaturation at 94C for 10 min, followed by 40 cycles of denaturation at 94C for 30 s, annealing at 55C for 30 s, and elongation at 72C for 2 min, followed by an elongation step at 72C for 30 min.