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Reverse Complement PCR: A novel one-step PCR system for typing highly degraded DNA for human identification

NCJ Number
302568
Journal
Forensic Science International-Genetics Volume: 44 Dated: 2020
Date Published
2020
Author(s)
R. E. Kieser; et al
Agencies
NIJ-Sponsored
Annotation

In this study, a human identification (HID) RC-PCR panel was designed to target 27 identity single nucleotide polymorphisms (SNPs) generating targets only 50 base pairs in length.

Abstract

Reverse Complement PCR (RC-PCR) is an innovative, one-step PCR target enrichment technology adapted for the amplification of highly degraded (fragmented) DNA. It provides simultaneous amplification and tagging of a targeted sequence construct in a single, closed-tube assay. In the current study, the complete sequencing construct was produced in a single reaction, which is essential for massively parallel sequencing (MPS) library preparation, thus reducing time and labor as well as minimizing the risk of sample carry-over or other forms of contamination. The RC-PCR system was evaluated and found to produce reliable and concordant variant calls. Also, the RC-PCR system demonstrated to have substantial sensitivity of detection with most alleles detected at 60 pg of input DNA and robustness in tolerating known PCR inhibitors. The RC-PCR system may be an effective alternative to current forensic genetic methods in the analysis of highly degraded DNA. (publisher abstract modified)

Date Created: September 25, 2021