Abstract
Analytical techniques such as DNA profiling are widely used in various fields, including forensic science, and novel technologies such as direct polymerase chain reaction (PCR) amplification are continuously being developed in order to acquire DNA profiles efficiently. However, non-specific amplification may occur depending on the quality of the crime scene evidence and amplification methods employed. In particular, the ski-slope effect observed in direct PCR amplification has led to inaccurate interpretations of the DNA profile results. In this study, we aimed to reduce the ski-slope effect by using dimethyl sulfoxide (DMSO) in direct PCR. We confirmed that DMSO (3.75%, v/v) increased the amplification yield of large-sized DNA sequences more than that of small-sized ones. Using 50 Korean buccal samples, we further demonstrated that DMSO reduced the ski-slope effect in direct PCR. These results suggest that the experimental method developed in this study is suitable for direct PCR and may help to successfully obtain DNA profiles from various types of evidence at crime scenes.
Highlights
In order to determine the appropriate concentration of dimethyl sulfoxide (DMSO) for the multiplex polymerase chain reaction (PCR) system, we first examined the amplification yield of 2800M
The amplification yield of 2800M DNA was observed to increase in a dose-dependent manner at high DMSO concentrations, and it was most effective at a concentration of 3.75% (Figure 1A)
We investigated the potential of DMSO in the reduction of ski-slope effect and enhancement of amplification in direct PCR system using Prep-n-Go buffer
Summary
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. First developed by Alec Jeffreys in 1985, DNA fingerprinting has evolved rapidly since the 1990s and found applications in domains such as criminal investigations, individual identification, and paternity tests [1,2,3]. The development of scientific investigation techniques leading to a high success rate of DNA extraction from human tissue samples—. Including crime scene samples—and improvements in short tandem repeat (STR) amplification have further resulted in more reliable DNA profiling results. More efficient direct polymerase chain reaction (PCR) amplification techniques for obtaining
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