Molecular Breeding

, Volume 3, Issue 5, pp 381–390

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

  • C.J. Jones
  • K.J. Edwards
  • S. Castaglione
  • M.O. Winfield
  • F. Sala
  • C. van de Wiel
  • G. Bredemeijer
  • B. Vosman
  • M. Matthes
  • A. Daly
  • R. Brettschneider
  • P. Bettini
  • M. Buiatti
  • E. Maestri
  • A. Malcevschi
  • N. Marmiroli
  • R. Aert
  • G. Volckaert
  • J. Rueda
  • R. Linacero
  • A. Vazquez
  • A. Karp
Article

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.

DNA markers RAPD AFLP SSR microsatellite network reproducibility 

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Copyright information

© Kluwer Academic Publishers 1997

Authors and Affiliations

  • C.J. Jones
    • 1
  • K.J. Edwards
    • 1
  • S. Castaglione
    • 2
  • M.O. Winfield
    • 2
  • F. Sala
    • 2
  • C. van de Wiel
    • 3
  • G. Bredemeijer
    • 3
  • B. Vosman
    • 3
  • M. Matthes
    • 4
  • A. Daly
    • 4
  • R. Brettschneider
    • 5
  • P. Bettini
    • 6
  • M. Buiatti
    • 6
  • E. Maestri
    • 7
  • A. Malcevschi
    • 7
  • N. Marmiroli
    • 7
  • R. Aert
    • 8
  • G. Volckaert
    • 8
  • J. Rueda
    • 9
  • R. Linacero
    • 9
  • A. Vazquez
    • 9
  • A. Karp
    • 1
  1. 1.IACR-Long Ashton Research Station, Department of Agricultural SciencesUniversity of BristolLong Ashton, BristolUK
  2. 2.Dipartimento di BiologiaUniversità di MilanoMilanoItaly
  3. 3.CPRO-DLOWageningenNetherlands
  4. 4.Jealott's Hill Research StationZeneca SeedsBracknell BerkshireUK
  5. 5.Institut für Allgemeine BotanikHamburgGermany
  6. 6.Dipartimento di Biologia Animale e GeneticaUniversità degli Studi di FirenzeFirenzeItaly
  7. 7.Dipartimento di Scienza AmbientaliUniversità degli Studi di ParmaParmaItaly
  8. 8.Laboratory of Gene TechnologyCatholic University of LeuvenLeuvenBelgium
  9. 9.Departamento de genetica, Facultad de BiologiaUniversidad ComplutenseMadridSpain

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