The methods and compositions provided herein relate to the discovery of several new STR alleles and a 20-plex multiplex assay for markers found on the human X chromosome. These X-STR markers have been found when used in a multiplex reaction to provide higher discriminatory power when compared to existing X-STR assay kits available commercially. Embodiments of the present teachings include methods for allelic determination of X-STR markers, amplification primers for the analysis of X-STR markers in a multiplex reaction, allelic ladders for analysis of X-STR markers, and kits for the analysis of X-STR markers.
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