Article

Molecular Breeding

, Volume 3, Issue 5, pp 381-390

First online:

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

  • C.J. JonesAffiliated withIACR-Long Ashton Research Station, Department of Agricultural Sciences, University of Bristol
  • , K.J. EdwardsAffiliated withIACR-Long Ashton Research Station, Department of Agricultural Sciences, University of Bristol
  • , S. CastaglioneAffiliated withDipartimento di Biologia, Università di Milano
  • , M.O. WinfieldAffiliated withDipartimento di Biologia, Università di Milano
  • , F. SalaAffiliated withDipartimento di Biologia, Università di Milano
  • , C. van de WielAffiliated withCPRO-DLO
  • , G. BredemeijerAffiliated withCPRO-DLO
  • , B. VosmanAffiliated withCPRO-DLO
  • , M. MatthesAffiliated withJealott's Hill Research Station, Zeneca Seeds
    • , A. DalyAffiliated withJealott's Hill Research Station, Zeneca Seeds
    • , R. BrettschneiderAffiliated withInstitut für Allgemeine Botanik
    • , P. BettiniAffiliated withDipartimento di Biologia Animale e Genetica, Università degli Studi di Firenze
    • , M. BuiattiAffiliated withDipartimento di Biologia Animale e Genetica, Università degli Studi di Firenze
    • , E. MaestriAffiliated withDipartimento di Scienza Ambientali, Università degli Studi di Parma
    • , A. MalcevschiAffiliated withDipartimento di Scienza Ambientali, Università degli Studi di Parma
    • , N. MarmiroliAffiliated withDipartimento di Scienza Ambientali, Università degli Studi di Parma
    • , R. AertAffiliated withLaboratory of Gene Technology, Catholic University of Leuven
    • , G. VolckaertAffiliated withLaboratory of Gene Technology, Catholic University of Leuven
    • , J. RuedaAffiliated withDepartamento de genetica, Facultad de Biologia, Universidad Complutense
    • , R. LinaceroAffiliated withDepartamento de genetica, Facultad de Biologia, Universidad Complutense
    • , A. VazquezAffiliated withDepartamento de genetica, Facultad de Biologia, Universidad Complutense
    • , A. KarpAffiliated withIACR-Long Ashton Research Station, Department of Agricultural Sciences, University of Bristol Email author 

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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.

DNA markers RAPD AFLP SSR microsatellite network reproducibility