Microsatellite Primer Amplification by Multiplexing: A First Application to Eucalyptus grandis

Abstract : Multiplexing was used to increase the efficiency of microsatellite analysis in Eucalyptus grandis. Primers were chosen based on the range of eucalyptus SSR alleles and their hybridization temperatures. Choice of PCR conditions proved to be the most impor- tant factor; concentrations of primers must be adjusted for use in combination. Results show that 6 primers can be analyzed in 2 PCR reactions, one containing 2 and the other containing 4 primers. Multiplexing promises to be a particularly efficient tool for micro- satellite diversity analysis in Eucalyptus grandis