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

TL;DR

This paper introduces a quick and reliable method to create degraded DNA for genetic testing and research purposes.

Contribution

The study presents a novel, rapid protocol for generating reproducibly degraded DNA using UV-C irradiation.

Findings

UV-C irradiation effectively reduces DNA fragment size in a reproducible manner within five minutes.

The degradation pattern is suitable for mimicking naturally degraded DNA in biological samples.

Degradation-sensitive PCR and STR analysis confirmed the method's reliability and reproducibility.

Abstract

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.

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