A non-destructive enzymatic method to extract DNA from arthropod specimens: Implications for morphological and molecular studies

Autor(es)

Santos, Daubian ^[1] ; Ribeiro, Guilherme Cunha ^[1] ; Cabral, Aline Diniz ^[1] ; Speranca, Marcia Aparecida ^[1]

Afiliação do(s) autor(es)

^[1] Univ Fed ABC, Ctr Ciencias Nat & Humanas, Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: PLoS One; v. 13, n. 2 FEB 1 2018.
• Citações Web of Science: 3

Resumo

There is a growing necessity to integrate morphological and genetic studies. This paper proposes a new technique that allows DNA extraction of arthropods while still keeping intact the entire morphology of the specimens. The technique uses Proteinase K to dissolve protein tissues and preserve the chitinous exoskeleton of specimens. The method is fast, cheap, non-toxic, and allows for good morphological preparations of specimens retaining much of their tridimensional structure. The methodology works fine with specimens preserved in different kinds of media, such as for dry (pinned) specimens, and specimens preserved in Ethanol. In addition, it allows the extraction of DNA from fresh specimens, as well as from specimens preserved for a long time. The technique works well for morphological studies alone, but allows the generation of an associated genomic library at an individual-scale. Among the advantages of the new technique is the possibility of extracting DNA from the entire specimen (necessary for the study of diseases transmitted by arthropod vectors), while still keeping the morphology intact for correct taxonomic identification. In addition, in comparison with methods that extract DNA from small tissue samples (e.g., from legs or wings), the method allows for the extraction of a larger amount of DNA and is better suited for small specimens. (AU)

• Processo FAPESP: 13/26096-4 - Expressão heteróloga da enzima quitinase de Leishmania (L.) infantum chagasi, Leishmania (V.) braziliensis e Leishmania amazonensis: diagnóstico sorológico e estudo molecular comparativo utilizando sistemas de expressão em células de insetos e bactéria
• Beneficiário: Aline Diniz Cabral
• Linha de fomento: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 16/14514-4 - Caracterização da quitinase das Espécies de Leishmania endêmicas da América do Sul: utilização no diagnóstico em humanos, cães e flebotomíneos
• Beneficiário: Marcia Aparecida Speranca
• Linha de fomento: Auxílio à Pesquisa - Regular