By modifying the cycling conditions and by combining the use of a DNA polymerase optimized for high speed PCR (SpeedSTAR HS) and a more efficient thermal cycler instrument (Bio-RAD C1000), the authors were able to reduce the amplification process from 4 h to 26 min. No modification to the commercial AmpFlSTR Profiler Plus primer mix was required. When compared to the current Royal Canadian Mounted Police (RCMP) amplification protocol, no differences with regards to specificity, sensitivity, heterozygote peak height ratios and overall profile balance were noted. Moreover, complete concordance was obtained with profiles previously generated with the standard amplification protocol and minor alleles in mixture samples were reliably typed. An increase in n - 4 stutter ratios (2.2 percent on average for all loci) was observed for profiles amplified with the fast protocol compared to the current procedure. The study results document the robustness of this rapid amplification protocol for STR profiling using the AmpFlSTR Profiler Plus primer set and demonstrate that comparable data can be obtained in substantially less time. This new approach could provide an alternative option to current multiplex STR typing amplification protocols in order to increase throughput or expedite time-sensitive cases. (Published Abstract)