A Rapid, Reliable and Reproducible Protocol for DNA Degradation in Genetic Applications

Degraded DNA is frequently encountered in biological samples due to mechanisms that favor the decomposition process and thus the fragmentation of DNA due to (extreme) environmental conditions. The reduced size of the DNA fragments may hamper the performance of genetic tests. This is why developmental evaluation and validation experiments of new markers and new technologies also involve the analysis of artificially degraded DNA. This study presents a method to reproducibly generate degraded DNA in only five minutes. Different concentrations and volumes of DNA extracted from blood were irradiated by UV-C light. This led to a gradual decrease in DNA fragment size that is typically targeted in genetic applications. An increase in DNA extract volume showed only little effect on the degradation performance; the starting DNA amount slightly shifted the observed degradation pattern. The process was assessed using degradation-sensitive quantitative real-time PCR and Short Tandem Repeat analysis. Repeated experiments resulted in comparable degradation patterns that were suitable for mimicking degradation states in biological samples and for evaluating genotyping applications.