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Design of dual-index sequencing strategy and schematic describing the four sequencing reads. The primers specific to the 16S rRNA gene are shown in boldface black text, linkers are in blue, pads are in green, the index region is in red, and the adapters are underlined. This schematic is demonstrated using the V4-specific primer sequences and linkers. The PCR and sequencing primers for each of the three regions are provided in the supplemental material.
