REPRODUCIBILITY TESTING OF RAPD, AFLP AND SSR MARKERS IN PLANTS BY A NETWORK OF EUROPEAN LABORATORIES

A number of PCR-based techniques can be used to detect polymorphisms i n plants. For their wide-scale usage in germplasm characterisation and breeding it is important that these marker technologies can be exchan ged between laboratories, which in turn requires that they can be stan dardised to yield reproducible results, so that direct collation and c omparison of the data are possible. This article describes a network e xperiment involving several European laboratories, in which the reprod ucibility of three popular molecular marker techniques was examined: r andom-amplified fragment length polymorphism (RAPD), amplified fragmen t length polymorphism (AFLP) and sequence-tagged microsatellites (SSR) . For each technique, an optimal system was chosen, which had been sta ndardised and routinely used by one laboratory. This system (genetic s creening package) was distributed to different participating laborator ies in the network and the results obtained compared with those of the original sender. Different experiences were gained in this exchange e xperiment with the different techniques. RAPDs proved difficult to rep roduce. For AFLPs, a single-band difference was observed in one track, whilst SSR alleles were amplified by all laboratories, but small diff erences in their sizing were obtained.