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.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Barbour T (1914) A contribution to the zoogeography of the West Indies, with special reference to amphibians and reptiles. Memoirs of the museum of comparative zoology. Cambridge, Massachusetts Vol 44, p 205–359
Bessetti J (2007) An introduction to PCR inhibitors. Profiles DNA 10:9–10
Egloff C, Lambrosse A, Hebert C, Crump D (2009) A nondestructive method for obtaining maternal DNA from avian eggshells and its application to embryonic viability determination in herring gulls (Larus argentatus). Mol Ecol Resour 9:19–27
Funk WC, Donnelly MA, Lips KR (2005) Alternative views of amphibian toe-clipping. Nature 433:193
Goldberg CS, Kaplan ME, Schwalbe CR (2003) From the frog’s mouth: buccal swabs for collection of DNA from amphibians. Herptol Rev 34:220–221
Gosner RA, Collura RV (1996) Waste not, want not: toe-clips as a source of DNA. J Herpetol 30:445–447
Hleap JS, Cárdenas H, García-Vallejo F (2009) Preservación no criogénica de tejido y extracción de DNA: una aplicación para peces cartilaginosos. PanamJAS 4:545–555
Kaiser H, Barrio-Amoros CL, Trujillo JD, Lynch JD (2002) Expansion of Eleutherodactylus johnstonei in northern South America. Herptol Rev 33:290–294
Kessing B, Croom H, Martin A, McIntosh C, Owen MW, and Palumbi SP (2004) The simple fool’s guide to PCR, version 1.0. Special publication. Dept. of Zoology, University of Hawaii, Honolulu, USA
Lopera-Barrero NM, Ribeiro RP, Sirol RN, Povh JA, Gomes PC, Streit DP Jr, Vargas LM (2008) Comparison of DNA extraction protocols of fish fin and larvae samples: modified salt (NaCl) extraction. Cienc Investig Agrar 35:65–74
Lucentini L, Palomba A, Lacioni H, Natali M, Fausto P (2006) A nondestructive, rapid, reliable and inexpensive method to sample, store and extract high-quality DNA from fish body mucus and buccal cells. Mol Ecol Notes 6:257–260
Mendoza AM, García-Ramírez JC, Cardenas-Henao H (2012) Blood puncture as a non-destructive sampling tool to obtain DNA in frogs: comparison of protocols and survival analysis. Mol Ecol Res 12:470–475
Parris KM, McCall SC, McCarthy MA, Minteer BA, Steele K, Bekessy S, Medvecky F (2010) Assessing ethical trade-offs in ecological field studies. J Appl Ecol 47:227–234
Perry G, Wallace MC, Perry D, Curzer H, Muhlberger P (2011) Toe clipping of amphibians and reptiles: science, ethics, and the law. J Herpetol 45:547–555
Pidancier N, Miquel NC, Miaud C (2003) Buccal swabs as a nondestructive tissue sampling method for DNA analysis in amphibians. Herpetol J 13:175–178
Poschadel JR, Möller D (2004) A versatile field method for tissue sampling on small reptiles and amphibians, applied to ponds turtles, newts, frogs and toads. Conserv Genet 5:865–867
Prunier J, Kaufmann B, Grolet O, Picard D, Pompanon F, Joly P (2012) Skin swabbing as a new efficient DNA sampling technique in amphibians, and 14 new microsatellite markers in the alpine newt (Ichthyosaura alpestris). Mol Ecol Res 12:524–553. doi:10.1111/j.1755-0998.2012.03116.x
Taberlet R, Walts LP, Luckart G (1999) Noninvasive genetic sampling: look before you leap. Trends Ecol Evol 14:323–327
Walsh PS, Metzger DA, Higuchi R (1991) Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. Biotechniques 10:506–513
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.
About this article
Cite this article
Mendoza, Á.M., García-Ramirez, J.C. & Cárdenas-Henao, H. Epithelial mucosa as an alternative tissue for DNA extraction in amphibians. Conservation Genet Resour 4, 1097–1099 (2012). https://doi.org/10.1007/s12686-012-9714-6
- Epithelial Mucosa
- Non-destructive sampling
- DNA extraction
- Comparison protocols