title: 16S RNA of field collected Amblyomma americanum ticks
design: Illumina dual indexes were used for library preparation.  All the primers for amplicon PCR (first PCR) and indexing PCR (second PCR) were ordered from Integrated DNA Technologies (IDT). Multiplexing of tick samples was performed by targeted 16S amplicon sequencing. The genespecific sequences used in this protocol target the 16S V3 and V4 regions. Illumina adapter overhang nucleotide sequences are added to the genespecific sequences. The full-length primer sequences for amplicon PCR (first PCR) were 5'-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCCTACGGGNGGCWGCAG-3 (Forward), and 5'-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG GACTACHVGGGTATCTAATCC-3 (Reverse) where forward primer overhang sequence is TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG, and reverse primer overhang sequence is GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG. V3-V4 locus-specific forward primer is CCTACGGGNGGCWGCAG, while reverse primer for the same is GACTACHVGGGTATCTAATCC.