A combination of techniques proves useful in the development of nuclear markers in the newt genus Triturus

  1. G. ESPREGUEIRA THEMUDO1,2,
  2. W. BABIK3,4,
  3. J. W. ARNTZEN1

Article first published online: 24 FEB 2009

DOI: 10.1111/j.1755-0998.2009.02574.x

Issue

Molecular Ecology Resources

Molecular Ecology Resources

Volume 9, Issue 3, pages 1160–1162, May 2009

Additional Information

How to Cite

ESPREGUEIRA THEMUDO, G., BABIK, W. and ARNTZEN, J. W. (2009), A combination of techniques proves useful in the development of nuclear markers in the newt genus Triturus. Molecular Ecology Resources, 9: 1160–1162. doi: 10.1111/j.1755-0998.2009.02574.x

Author Information

  1. 1

    National Museum of Natural History Naturalis, PO Box 9517, 2300 RA Leiden, The Netherlands,

  2. 2

    CIBIO, Centro de Investigação em Biodiversidade e Recursos Genéticos, Campus Agrário de Vairão, 4485-661 Vairão, Portugal,

  3. 3

    Department of Community Ecology, Helmholtz Centre for Environmental Research — UFZ, Theodor-Lieser-Strasse 4, 06120 Halle (Saale), Germany,

  4. 4

    Institute of Environmental Research, Jagiellonian University, Gronostajowa 7, 30-387 Krakow, Poland

* Gonçalo Espregueira Themudo, Fax: +31 71 568 7666; E-mail: themudo@nnm.nl

Publication History

  1. Issue published online: 20 APR 2009
  2. Article first published online: 24 FEB 2009
  3. Received 21 October 2008; revision accepted 11 December 2008

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Keywords:

  • Amphibia;
  • anonymous markers;
  • introns;
  • newts;
  • nuclear DNA markers;
  • Triturus

Abstract

To increase the number of markers available for study of phylogeny and phylogeography in the newt genus Triturus, we developed and tested 59 primer pairs using three different techniques. Primers were obtained from published sources, by designing exon-primed intron-crossing primers and from randomly cloned anonymous nuclear DNA fragments. Successful polymerase chain reaction products were cloned and sequenced. Five fragments were successfully amplified and sequenced for six species of Triturus: intron 7 of the β-fibrinogen gene (βfibint7), third intron of the calreticulin gene (CalintC), the 11th intron of the α-subunit of the platelet derived growth factor receptor (PDGFRα) and two anonymous markers (Cri1 and Cri4). The average percentage species divergence across all the markers is low (c. 3%), compared to what has been found in mitochondrial DNA (25–30%).

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