Reproducibility Testing of RAPDs by a Network of European Laboratories

Abstract

For the application of molecular techniques to wide-scale diversity screening it is important that the different screening techniques employed can be standardized to yield reproducible results across laboratories in order that direct collation and comparison of the data are possible (1). RAPDs (Random Amplified Polymorphic DNA) involve the use of a single ‘arbitrary’ primer (purchasable from commercial companies) in a PCR reaction and results in the amplification of several discrete DNA products (see Chapter 9). It is now widely recognized that to obtain reproducible band profiles on the gels it is absolutely essential to maintain consistent reaction conditions. Numerous studies have reported the separate effects of altering different parameters, such as ratio of template DNA/primers, concentration of Taq polymerase and Mg concentration, on the bands obtained (2–5). A network reproducibility test was carried by nine European laboratories (L1-L9) to determine whether RAPD profiles could be reproduced by different laboratories if all details of the reaction conditions were standardized (1).