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

A - Papers appearing in refereed journals

Jones, C. J., Edwards, K. J., Castaglione, S., Winfield, M. O., Sala, F., van de Wiel, C., Bredemeijer, G., Vosman, B., Matthes, M., Daly, A., Brettschneider, R., Bettini, P., Buiatti, M., Maestri, E., Malcevschi, A., Marmiroli, N., Aert, R., Volckaert, G., Rueda, J., Linacero, R., Vazquez, A. and Karp, A. 1997. Reproducibility testing of RAPD, AFLP and SSR markers in plants by a network of European laboratories. Molecular Breeding. 3 (5), pp. 381-390.

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

KeywordsDNA markers; Rapd; AFLP; SSR; microsatellite; network; reproducibility; polymorphic DNA markers; arbitrary primers; Pcr; Identification; Microsatellites; Cultivars; Rflp; Loci
Year of Publication1997
JournalMolecular Breeding
Journal citation3 (5), pp. 381-390
Digital Object Identifier (DOI)doi:10.1023/a:1009612517139
Open accessPublished as non-open access
FunderBiotechnology and Biological Sciences Research Council
ISSN13803743

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