The aim of the present study was to determine a supplementary set of microsatellite DNA markers to be used in addition to the standard panel of 12 microsatellite markers recommended for parentage testing in cattle. A method has been developed for analysis of 10 loci in a single multiplex PCR reaction and using automated DNA sizing technology in a sequencer, which will enable the additional set of markers to be used for routine pedigree verification. Preliminary analysis showed that the polymorphism of 10 selected markers (PIC and H > 0.5 for 9 loci) and PEc of 99.77% suggest that the microsatellite sequences selected could be useful in cases of disputable parentage where the standard set may prove insufficient, e.g. in closely related animals or those from herds subjected to rigorous selection.