Conservation Genetics Resources

, Volume 4, Issue 4, pp 1097–1099 | Cite as

Epithelial mucosa as an alternative tissue for DNA extraction in amphibians

  • Ángela María Mendoza
  • Juan Carlos García-Ramirez
  • Heiber Cárdenas-Henao
Technical Note

Abstract

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.

Keywords

Epithelial Mucosa Non-destructive sampling Amphibians DNA extraction mtDNA Comparison protocols 

References

  1. 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 205–359Google Scholar
  2. Bessetti J (2007) An introduction to PCR inhibitors. Profiles DNA 10:9–10Google Scholar
  3. 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 9:19–27PubMedCrossRefGoogle Scholar
  4. Funk WC, Donnelly MA, Lips KR (2005) Alternative views of amphibian toe-clipping. Nature 433:193PubMedCrossRefGoogle Scholar
  5. Goldberg CS, Kaplan ME, Schwalbe CR (2003) From the frog’s mouth: buccal swabs for collection of DNA from amphibians. Herptol Rev 34:220–221Google Scholar
  6. Gosner RA, Collura RV (1996) Waste not, want not: toe-clips as a source of DNA. J Herpetol 30:445–447CrossRefGoogle Scholar
  7. Hleap JS, Cárdenas H, García-Vallejo F (2009) Preservación no criogénica de tejido y extracción de DNA: una aplicación para peces cartilaginosos. PanamJAS 4:545–555Google Scholar
  8. Kaiser H, Barrio-Amoros CL, Trujillo JD, Lynch JD (2002) Expansion of Eleutherodactylus johnstonei in northern South America. Herptol Rev 33:290–294Google Scholar
  9. 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, USAGoogle Scholar
  10. 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 35:65–74Google Scholar
  11. 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 6:257–260CrossRefGoogle Scholar
  12. 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 12:470–475CrossRefGoogle Scholar
  13. 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 47:227–234CrossRefGoogle Scholar
  14. Perry G, Wallace MC, Perry D, Curzer H, Muhlberger P (2011) Toe clipping of amphibians and reptiles: science, ethics, and the law. J Herpetol 45:547–555CrossRefGoogle Scholar
  15. Pidancier N, Miquel NC, Miaud C (2003) Buccal swabs as a nondestructive tissue sampling method for DNA analysis in amphibians. Herpetol J 13:175–178Google Scholar
  16. 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 5:865–867Google Scholar
  17. 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 CrossRefGoogle Scholar
  18. Taberlet R, Walts LP, Luckart G (1999) Noninvasive genetic sampling: look before you leap. Trends Ecol Evol 14:323–327PubMedCrossRefGoogle Scholar
  19. 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 10:506–513PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Ángela María Mendoza
    • 1
  • Juan Carlos García-Ramirez
    • 2
  • Heiber Cárdenas-Henao
    • 3
  1. 1.Laboratorio de Macroecología, Centro de Investigaciones en ecosistemasUniversidad Nacional Autónoma de MéxicoMoreliaMexico
  2. 2.Institute of Natural ResourcesMassey UniversityPalmerston NorthNew Zealand
  3. 3.Grupo de Estudios Ecogenéticos y de Biología Molecular, Departamento de BiologíaUniversidad del ValleCaliColombia

Personalised recommendations