Cutaneous leishmaniasis induces a transmissible dysbiotic skin microbiota that promotes skin inflammation
Bacterial 16S rDNA microbiome sequencing of the V4 variable region
The V4 region of the 16S rRNA gene was amplified using forward primer 27F and reverse primer 534R and the Accumprime kit. Primers contained unique error-correcting barcodes to facilitate multiplexing of samples. Due to the heterogeneous template mixture, PCR amplification was performed in duplicate and resulting products pooled to reduce amplification bias. PCR products were purified using Agencour AMPure kit, normalized, and then subject to Illumina MiSeq paired-end sequencing. See Gardner SE, et.al, Diabetes. 2013 62(3):923–30 for details