Methods for the recovery of mitochondrial DNA sequences from museum specimens of myiasis-causing flies.
Mitochondrial DNA (mtDNA) sequences from eight species of myiasis-causing flies, stored for up to 50 years, were amplified successfully. Universal primers were used to amplify six specific regions from total genomic DNA, including five mtDNA genes. The comparison of phenol/chloroform, DNAzol and Chelex techniques for DNA extraction showed that the DNAzol reagent was the most efficient in retrieving DNA from museum specimens, although the Chelex extraction procedure is currently the most frequently reported method. Comparison of the universal primer sequences with the homologous sequences of Cochliomyia hominivorax Coquerel and Chrysomya putoria Wiedemann (Diptera: Calliphoridae) revealed mismatches that could contribute to the low recovery of a short sequence from subunit II of cytochrome oxidase. The ability to characterize mtDNA markers from museum specimens should be useful in comparative studies of contemporary samples and should help in elucidating species introduction, colonization and dispersal.
Junqueira
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AC
Laboratório de Genética Animal, Universidade Estadual de Campinas (UNICAMP), São Paulo, Brazil. anacmj@obelix.unicamp.br
Lessinger
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AC
Azeredo-Espin
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AM
D000818:Animals; D001483:Base Sequence; D001938:Brazil; D004272:DNA, Mitochondrial / Q000737:chemistry*; D004175:Diptera / Q000235:genetics*; D003576:Electron Transport Complex IV / Q000737:chemistry / Q000235:genetics; D005260:Female; D005784:Gene Amplification; D008297:Male; D008969:Molecular Sequence Data; D009198:Myiasis / Q000469:parasitology*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D017385:Sequence Homology
D004272:DNA, Mitochondrial; D003576:Electron Transport Complex IV