In seeking improvements in DNA typing methods on aspects such as sensitivity and efficacy, this study examined the effects of reduced volume PCR amplification (RV-PCR) through a series of five experiments. The five experiments consisted of (1) reducing proportionally to the amplification reaction volume the quantity of template DNA, (2) reducing the reaction volume while holding the amount of template DNA consistent, (3) determining the efficiency of the PCR amplification at reduced reaction volumes, (4) determining the minimum detection (sensitivity) for the experimental volumes, and (5) evaluating the ability to identify and interpret mixed samples at reduced reaction volumes. The DNA for the experiments was obtained from dried bloodstain cards and buccal swabs. Results indicated that reducing the PCR reaction volume below the recommended 50uL volume increased the sensitivity of the AmpFlSTR Profiler Plus reaction. It was found that smaller amounts of DNA could be amplified to the same concentration in reduced volume reactions. Three experiments clearly demonstrated that reducing the reaction volume increased the sensitivity which was proportional to the volume reduction. This examination of the benefits and pitfalls of reduced volume PCR reactions showed that these reactions could be useful in forensic analyses where extremely sensitive methods are required. References