DNA barcoding using chitons (genus Mopalia )

Incorporating substantial intraspecific genetic variation for 19 species from 131 individual chitons, genus Mopalia (Mollusca: Polyplacophora), we present rigorous DNA barcodes for this genus as per the currently accepted approaches to DNA barcoding. We also have performed a second kind of analysis that does not rely on blast or the distance‐based neighbour‐joining approach as currently resides on the Barcode of Life Data Systems website. Our character‐based approach, called characteristic attribute organization system, returns fast, accurate, character‐based diagnostics and can unambiguously distinguish between even closely related species based on these diagnostics. Using statistical subsampling approaches with our original data matrix, we show that the method outperforms blast and is equally effective as the neighbour‐joining approach. Our approach differs from the neighbour‐joining approach in that the end‐product is a list of diagnostic nucleotide positions that can be used in descriptions of species. In addition, the diagnostics obtained from this character‐based approach can be used to design oligonucleotides for detection arrays, polymerase chain reaction drop off diagnostics, Taq Man assays, and design of primers for generating short fragments that encompass regions containing diagnostics in the cytochrome oxidase I gene.