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Analysis of 12 X- chromosome STR LOCI with reduced PCR sample volume

STR markers on the X chromosome have emerged as additional tools in forensics, in missing person cases and parent-child relationships. Qiagen's Investigator® Argus X-12 kit for X- chromosome STR's, which analyses 12 loci (DXS10103, DXS8378, DXS7132, DXS10134, DXS10074, DXS10101, DXS10135, DXS7423, DXS10146, DXS10079, HPRTB, DXS10148) and amelogenin is currently being used. Following manufactures analysis protocol, obtained results from female samples were exhibiting spikes and pull- ups in electrophoresis, which were not found in male samples. Since the standard procedure requires samples to be normalized to concentrations of 1ng/µL of DNA before conducting PCR amplification, results from both male and female samples should be coherent. Due to observed spikes and pull-ups in female samples we diluted female and male normalized samples to 0.5ng/µL of DNA before setting up PCR reaction. Obtained results with diluted samples showed no spikes and pull-ups in female DNA profiles, while allele drop- out was observed in male DNA profiles. Thus we recommend normalization of female samples s to 0.5ng/µL of DNA rather than 1ng/µL of DNA asproposed by manufacturer.

Forensics ; DNA analysis ; Investigator Argus X-12 ; X chromosome ; Genetics

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