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An efficient and reproducible protocol for production of AFLP markers in tree genomes using fluorescent capillary detection

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Abstract

An optimized protocol for the development and discovery of polymorphic AFLP markers in tree species is described. The protocol was optimized for the production of fluorescently labeled PCR products and analysis using a capillary sequencer. This approach has been demonstrated to be efficient and reproducible for tree species with complex genomes. The most important modification was in the selective amplification step. Instead of using a traditional step down PCR, a fixed and higher annealing temperature was employed, improving the reproducibility and sensitivity of the protocol. The levels of polymorphisms detected with the optimized protocol on three woody species are in agreement with those previously reported in the literature for tree species.

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Acknowledgments

The authors gratefully acknowledge the support from INNOVA-CHILE (grant number 05CTE04-03) and Genómica Forestal. Ross Whetten is gratefully acknowledged for the critical reading of this manuscript and discussions throughout this work.

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Authors and Affiliations

  1. Centro de Biotecnología, Genómica Forestal S.A., Universidad de Concepción, Casilla 160 C, Concepción, Chile

    Rodrigo Hasbún, Carolina Iturra & Pamela Wachtendorff

  2. Facultad de Ciencias Forestales, Universidad de Concepción, Casilla 160 C, Concepción, Chile

    Rodrigo Hasbún, Priscila Moraga, Pamela Quiroga & Sofía Valenzuela

  3. Centro de Biotecnología, Universidad de Concepción, Victoria 631, Casilla 160 C, Concepción, Chile

    Sofía Valenzuela

Authors
  1. Rodrigo Hasbún
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  2. Carolina Iturra
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  3. Priscila Moraga
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  4. Pamela Wachtendorff
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  5. Pamela Quiroga
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  6. Sofía Valenzuela
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Corresponding author

Correspondence to Rodrigo Hasbún.

Additional information

Communicated by S. González-Martínez

The AFLP process is covered by patents and patent applications held by Keygene NV (Wageningen, Netherlands), and users of the technique should either purchase a commercial kit that confers a license to use the technique or contact Keygene directly regarding licensing.

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Hasbún, R., Iturra, C., Moraga, P. et al. An efficient and reproducible protocol for production of AFLP markers in tree genomes using fluorescent capillary detection. Tree Genetics & Genomes 8, 925–931 (2012). https://doi.org/10.1007/s11295-011-0463-6

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  • DOI: https://doi.org/10.1007/s11295-011-0463-6

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