16S rDNA of stool from Haitian adults
Primers used to amplify rDNA were: 563 F (59-nnnnnnnn-NNNNNNNNNNNN-AYTGGGYDTAAAGN G-39) and 926 R (59-nnnnnnnn-NNNNNNNNNNNN-CCGTCAATTYHTTTR AGT-39). Each reaction contained 50 ng of purified DNA, 0.2 mM dNTPs, 1.5  M MgCl2, 1.25 U Platinum TaqDNA polymerase, 2.5  l of 10   PCR buffer and 0.2  M of each primer. A unique 12-base Golay barcode (Ns) preceded the primers for sample identification after pooling amplicons. One to eight additional nucleotides were added before the barcode to offset the sequencing of the primers. Cycling conditions were the following: 94  C for 3 min, followed by 27 cycles of 94  C for 50 s, 51  C for 30 s and 72  C for 1 min, where the final elongation step was performed at 72  C for 5 min. doi:10.1038/s41598-017-10346-6