Analytical improvements that can achieve a reduction in evidential sample consumption in forensic DNA typing are constantly sought in forensic biology. The consumption of an entire sample of evidence in DNA analysis is logistically undesirable because it precludes further analysis. One way to achieve a reduction in evidential sample consumption involves the concomitant reduction of both the amount of template DNA and PCR reaction volume, so as to maintain the same DNA concentration and yield per unit volume. The impact of volume reduction was initially assessed on a collection of pristine single-source samples used in prepared mixtures. Additionally, forensically relevant and challenged samples from adjudicated cases were also analyzed. All samples were assessed under various PCR reaction volume/amount of input template DNA ratios and subjected to amplification with the AmpFlSTRProfiler Plus kit. The profiles were assessed for signal strength and quality consistency. The reduction of PCR reaction volume and DNA down to 10 ml and 0.063 ng yielded HR values that were slightly affected in one to three STR loci. The study concluded that PCR reaction volume reduction can enhance detection and sensitivity while reducing the consumption of irreplaceable crime-scene samples of evidence. 7 figures and 29 references