Optimisation of a multiplex enPolymerase Chain ReactionPCR assay of nuclear microsatellite markers for population genetics and clone identification in Robinia pseudoacaciaL.

Black locust (Robinia pseudoacacia L.) is a tree species native to North America. The multipurpose tree is cultivated worldwide, but causes problems due to its partially invasive character. The application of nuclear microsatellite loci has many aims in population genetic studies. Here we introduce a very cost-effective method for combining the information of 14 nuclear microsatellite loci into two multiplex PCR sets as a contribution to greater standardisation and more comparable results. Combined non-exclusion probabilities for clone identification using example populations are estimated at between 1.37*E-5 and 1.67*E-11, and for paternity analysis for 1.59*E-4. The detected weak linkage between some microsatellite loci is not considered to be a substantial restriction to the reliability of the set of markers in providing an appropriate method for fingerprinting and parentage analysis.