One of the first questions asked about excavated human skeletal remains is the 
sex. As the morphological sex determination is complicated in cases involving 
fragmentary bones and in skeletons from infants and children, the development of 
DNA-based techniques has led to improvements in sex determination. This study is 
focused on sex determination from ancient DNA obtained from 25 skeletons found 
in Middle Aged burials in western Slovakia. We performed separate amplifications 
of DXZ4 repetitive satellite sequences on the X chromosome, and SRY gene - 
testis determined factor on the Y chromosome, using nested PCR. Our results 
showed that DXZ4 was amplified in the case of 23 individuals. With newly 
designed internal and external primer sets for SRY detection with internal PCR 
products in lengths of 102 bp and 85 bp we succeeded in detecting the SRY locus 
in 9 samples. Finally, the gender was determined in 23 individuals (14 females 
and 9 males). In 20 samples, the gender was determined by morphological and 
molecular methods. Sex determination of 17 samples using nested PCR matched the 
morphological one, providing evidence of the authenticity and ancient origin of 
the PCR amplifications. The DXZ4/SRY nested PCR method represents a useful 
technique in sex determination of medieval human remains and it is a critical 
addition to anthropological studies.