A small and inexpensive thermal cycler (PCR machine), known as the MiniPCR TM Mini8 Thermal Cycler (Amplyus, Cambridge, MA, USA), was developed. In this study, the performance of this PCR machine was compared with the GeneAmp ® PCR system 9700 (Applied Biosystems) using four autosomal short tandem repeat (STR) kits, a Y-chromosome STR kit, and a mitochondrial DNA HV1/HV2 sequence analysis. The sensitivity and stochastic effects of the STR multiplex kits and the quality of the DNA sequence analysis were similar between the two PCR machines. The MiniPCR TM Mini8 Thermal Cycler could be used for analyses at forensic DNA laboratories and crime scenes. The cost of the PCR is so economical that school laboratories and individuals could use the machines.
The Quantifiler Trio Quantification Kit has been developed to quantify the total amount of amplifiable and human male DNA in samples and to estimate the extent of DNA degradation. To minimize the cost of DNA quantification, we evaluated kit performance using a reduced volume of reagents (1/10-volume) using DNA samples of varying types and concentrations. Our results demonstrated concordance between the manufacturer's method and the low-volume method for DNA quantification, DNA degradation index estimation, and human male DNA quantification. We confirmed the practical utility of the low-volume method with 109 casework samples by evaluating short tandem repeat (STR) profiling success with respect to DNA quantity and quality. We also defined a cutoff value for DNA quantity to ensure reliable STR results. Using a reduced volume of reagents, 10 times more reactions per kit are possible; accordingly, this method reduces the cost of DNA quantification, while maintaining performance.
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