This article describes the results of a study to develop a quantitative PCR assay for the assessment of DNA degradation in forensic samples.
The study found that for highly degraded DNA samples, the rate of genotyping success improved when an appropriately long qPCR target was used for determining the amount of template needed for amplification for STR (short tandem repeat) analysis. The study also found that a degradation ratio was able to be calculated from quantitation results obtained with the nuTH01-nuCSF-IPC triplex qPCR assay, and that this ratio proved to be a good estimate of the quality of the DNA extracted from the degraded sample. The primary objective of the study was to develop a quantitative PCR (qPCR) assay for use in assessing the degradation of DNA in forensic samples. Data for the study were obtained from analyses of both non-degraded and highly degraded DNA samples. The nuTH01 and nuCSF qPCR assays were tested on the samples in order to ensure that equivalent results could be obtained in both singleplex and duplex reactions. The IPC assay was then added to the nuTH01-nuCSF duplex assay. This triplex assay was then tested on the DNA samples and found to be highly successful at quantifying DNA in the degraded samples. This finding indicates that the nutH01-nuCSF-IPC triplex assay can be a useful tool for simultaneously assessing the quantity and quality of DNA, thereby minimizing the need for large of amounts of extracted DNA and maximizing the throughput and efficiency of DNA analysis. Tables, figures, and references