This paper presents the methodology behind the development of a Reverse Complement PCR system that may be an effective alternative to current forensic genetic methods in the analysis of highly degraded DNA.
Compromised biological samples are one of the main challenges facing forensic analysts because they are difficult to genetically profile, due to the highly fragmented nature of the target molecules. To address this issue, the paper presents the use of an innovative, one-step PCR target enrichment technology that was adapted for the amplification of highly degraded, or fragmented, DNA: Reverse Complement PCR (RC-PCR), which provides simultaneous amplification and tagging of a targeted sequence construct in a single, closed-tube assay. Results indicated that RC-PCR provided reliable and concordant genotyping results as well as robustness in tolerating known PCR inhibitors such as calcium and collagen, suggesting that the system may be an effective alternative to current forensic genetic methods for analyzing highly degraded DNA.
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