16S rRNA of Campylobacter infection Colombia: child feces
Total microbial DNA was isolated from weighed, archival feces using the DNeasy PowerSoil Kit (QIAGEN) following the manufacturers protocol. DNA was stored at -20C until 16S rRNA gene amplification. Primers targeting the V3-4 region of the 16S rRNA gene were used to create a single amplicon of approximately 460 bp: 16S V3-4 Amplicon PCR Forward Primer (5'-CCTACGGGNGGCWGCAG-3) and 16S V3-4 Amplicon PCR Reverse Primer (5'-GACTACHVGGGTATCTAATCC-3) according to the Illumina 16S Metagenomic Sequencing Library Preparation Guide. Once the library was quantified and normalized, it was pooled and sequenced on an Illumina MiSeq using v3 600 cycle kit (2 x 300 bp).