Evaluation location: Puerto Rico, United States
Fifteen microsatelite markers were used to fingerprint all 154 accessions of the current USDA-ARS TARS cacao germplasm collection. Fingerprint profiles generated were used to determine inter-clonal and intra-clonal error, as well as identifying potential genetic gaps and estimating genetic diversity among current accessions. A Fast DNA SPIN Kit (MP Biomedicals, Irvine, CA) was utilized for DNA extraction using the procedure described in the manual with minor modifications. Forward primers were labeled with one of three fluorescent dyes on the 5' end and all 15 primer pairs were used on all individuals for the analysis in a 10 ul reaction. Capillary electrophoresis was performed on an ABI Prism 3730 Genetic Analyzer. Electrophoresis results were analyzed with GeneMapper 3.0 software to determine alleles and for internal standard and fragment size determination.