Epithelial mucosa as an alternative tissue for DNA extraction in amphibians
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We evaluated the performance of amphibian epithelial mucosa as a non-destructive method for sampling DNA in four extraction protocols. We took tissue from 68 individuals of Eleutherodactylus johnstonei (Anura: Eleutherodactylidae) through a surface smear of each specimen with a sterile swab. DNA was extracted using the DNeasy extraction kit (Qiagen), Salting-out, Phenol–chloroform, and Chelex protocols. We compared the quality of the resulting DNA through amplification and sequencing of the 16S rRNA mitochondrial gene. Successful amplification was obtained from DNA isolated from two protocols (Salting out and the DNeasy kit). The resulting sequences corresponded to those registered in the GenBank for this species, demonstrating that epithelial mucosa it is a valuable alternative method for obtaining DNA in frogs.
KeywordsEpithelial Mucosa Non-destructive sampling Amphibians DNA extraction mtDNA Comparison protocols
This project was funded by the COLCIENCIAS Program for Young Researchers and Innovators, Universidad del Valle, Group for Ecogenetic Studies and Molecular Biology and the Humboldt Institute. We thank to the Group for Studies on Reptiles and Amphibians (GERA), SAMANEA foundation, Valentina Rodríguez and María Isabel Arce for their help with field and laboratory work; Sergio Hleap for his comments on the experimental design and final manuscript; two anonymous reviewers for their helpful comments on a previous version of the manuscript; Vicerrectoría de Investigaciones (Universidad del Valle, Colombia) Andrew J. Crawford (Universidad de los Andes, Colombia) and Rachel Turner (Massey University, New Zealand) for revision of the English.
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