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Reproducibility testing of RAPD, AFLP and SSR markers in plants by a network of European laboratories

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Abstract

A number of PCR-based techniques can be used to detect polymorphisms in plants. For their wide-scale usage in germplasm characterisation and breeding it is important that these marker technologies can be exchanged between laboratories, which in turn requires that they can be standardised to yield reproducible results, so that direct collation and comparison of the data are possible. This article describes a network experiment involving several European laboratories, in which the reproducibility of three popular molecular marker techniques was examined: random-amplified fragment length polymorphism (RAPD), amplified fragment length polymorphism (AFLP) and sequence-tagged microsatellites (SSR). For each technique, an optimal system was chosen, which had been standardised and routinely used by one laboratory. This system (genetic screening package) was distributed to different participating laboratories in the network and the results obtained compared with those of the original sender. Different experiences were gained in this exchange experiment with the different techniques. RAPDs proved difficult to reproduce. For AFLPs, a single-band difference was observed in one track, whilst SSR alleles were amplified by all laboratories, but small differences in their sizing were obtained.

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Authors and Affiliations

  1. IACR-Long Ashton Research Station, Department of Agricultural Sciences, University of Bristol, Long Ashton, Bristol, BS18 9AF, UK

    C.J. Jones, K.J. Edwards & A. Karp

  2. Dipartimento di Biologia, Università di Milano, 20133, Milano, Italy

    S. Castaglione, M.O. Winfield & F. Sala

  3. CPRO-DLO, PO Box 16, 6700 AA, Wageningen, Netherlands

    C. van de Wiel, G. Bredemeijer & B. Vosman

  4. Jealott's Hill Research Station, Zeneca Seeds, Bracknell Berkshire, RG12 6EY, UK

    M. Matthes & A. Daly

  5. Institut für Allgemeine Botanik, Ohnhorstrasse 18, 22609, Hamburg, Germany

    R. Brettschneider

  6. Dipartimento di Biologia Animale e Genetica, Università degli Studi di Firenze, 17, Via Romana, 50125, Firenze, Italy

    P. Bettini & M. Buiatti

  7. Dipartimento di Scienza Ambientali, Università degli Studi di Parma, Via delle Scienze, 43100, Parma, Italy

    E. Maestri, A. Malcevschi & N. Marmiroli

  8. Laboratory of Gene Technology, Catholic University of Leuven, Willem de Croylaan 42, Leuven, 3001, Belgium

    R. Aert & G. Volckaert

  9. Departamento de genetica, Facultad de Biologia, Universidad Complutense, Ciudad Universitaria, Madrid, 28040, Spain

    J. Rueda, R. Linacero & A. Vazquez

Authors
  1. C.J. Jones
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  2. K.J. Edwards
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  3. S. Castaglione
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  4. M.O. Winfield
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  5. F. Sala
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  6. C. van de Wiel
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  7. G. Bredemeijer
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  8. B. Vosman
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  9. M. Matthes
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  10. A. Daly
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  11. R. Brettschneider
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  12. P. Bettini
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  13. M. Buiatti
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  14. E. Maestri
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  15. A. Malcevschi
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  16. N. Marmiroli
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  17. R. Aert
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  18. G. Volckaert
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  19. J. Rueda
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  20. R. Linacero
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  21. A. Vazquez
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  22. A. Karp
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Corresponding author

Correspondence to A. Karp.

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Jones, C., Edwards, K., Castaglione, S. et al. Reproducibility testing of RAPD, AFLP and SSR markers in plants by a network of European laboratories. Molecular Breeding 3, 381–390 (1997). https://doi.org/10.1023/A:1009612517139

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  • DOI: https://doi.org/10.1023/A:1009612517139

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