doi: 10.1371/journal.pone.0013716ĭoublet V, Raimond R, Grandjean F, Lafitte A, Souty-Grosset C, Marcadé I (2012) Widespread atypical mitochondrial DNA structure in isopods (Crustacea, Peracarida) related to a constitutive heteroplasmy in terrestrial species. Nature 378:485–489ĭerycke S, Vanaverbeke J, Rigaux A, Backeljau T, Moens T (2010) Exploring the use of cytochrome oxidase c subunit 1 (COI) for DNA barcoding of free-living marine nematodes. doi: 10.3732/ajb.Ĭollura RV, Stewart CB (1995) Insertions and duplications of mtDNA in the nuclear genomes of Old World monkeys and hominoids. Int Microbiol 1:327–332Ĭhristianson ML (2005) Codon usage patterns distort phylogenies from or of DNA sequences. Hydrobiologia 401:239–254Ĭastro JA, Picornell A, Ramon M (1998) Mitochondrial DNA: a tool for populational genetics studies. doi: 10.1007/s0043-8īucklin A, Guarnieri M, Hill RS, Bentley AM, Kaartvedt S (1999) Taxonomic and systematic assessment of planktonic copepods using mitochondrial COI sequence variation and competitive, species-specific PCR. Mol Ecol 20:503–516īoehme P, Amendt J, Zehner R (2012) The use of COI barcodes for molecular identification of forensically important fly species in Germany. doi: 10.1111/j.īock DG, Zhan A, Lejeusne C, MacIsaac HJ, Cristescu ME (2011) Looking at both sides of the invasion: patterns of colonization in the violet tunicate Botrylloides violaceus. doi: 10.1899/10-013.1īlankenship LE, Yayanos AA (2005) Universal primers and PCR of gut contents to study marine invertebrate diets. Reverse primers can be designed from the level of family (maximum 5 bp differences) or genus (maximum 2 bp differences).īaird DJ, Pascoe TJ, Zhou X, Hajibabaei M (2011) Building freshwater macroinvertebrate DNA-barcode libraries from reference collection material: formalin preservation vs specimen age. We conclude that, at different taxonomic levels, it is possible to design forward primers from reference sequences belonging to the level of order (maximum 5 bp differences), family (maximum 6 bp differences) or genus (maximum 1 bp difference). There was a significant difference in the means of base pair differences from the level of family, genus and species for LCO 1490 and at the level of genus and species for HCO 2198 (ANOVA, F 6,77 = 2.538, P < 0.027). A similar pattern was observed in amino acid distributions. However, the reverse primer (HCO 2198) was highly conserved across 725 COI primer sequences. For ascidians, fungi and vertebrates, it showed approximately 50 % conserved regions, dropping to one conserved region for echinoderms. It was found that, for 177 invertebrate species, the forward primer (LCO 1490) showed only four conserved regions, compared to 12 in the original study. In the present study, 130,843 variations were reviewed in the primer region of mitochondrial molecular markers by comparing 725 COI sequences from the kingdom Animalia. These primers have been successful in amplifying a 710-bp fragment of highly conserved regions of the COI gene for more than 80 invertebrate species from 11 phyla. “Universal” DNA primers LCO 1490 and HCO 2198 were originally designed from three coding and six anticoding strands by comparing highly conserved regions of mitochondrial cytochrome c oxidase subunit I ( COI) genes across 15 taxa.