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Information on genetic variation of breeding materials helps maintain genetic diversity and sustains long term selection gain. Marama bean [Tylosema esculentum (Burchell) Schreiber] occurs naturally in arid, dry parts of Southern Africa and due to the high nutrient value of the seeds and tubers, rich in protein, oil and starch, it is a potential crop for arid areas where few conventional crops can survive. Recently, the development of SSR enrichment techniques has increased the efficiency of SSR characterization in new species. The microsatellite regions of the genome were the main focus and the matK gene was also explored for its potential to carry out a marama bean genetic diversity study. Microsatellite loci were isolated from the marama bean germplasm using a modified FIASCO enrichment technique. Nine marama bean microsatellite libraries enriched for (AAG)7, (GTT)7, (AGG)7, (GAG)7, (CA)10, (CT)10, (TCC)7, (CA)15 and (CAC)7 were created. The matK gene product was found to be half the expected size of the gene found in other legumes. The found homology with Tylosema fassoglensis (trnK gene) and Pisum sativum (matK gene) suggests that an identical region was amplified.