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Application study of PCR additives to improve the split peaks in direct PCR

분석과학 2019년 32권 4호 p.155 ~ 162
 ( Kim Joo-Young ) - National Forensic Service Forensic DNA Division

김다혜 ( Kim Da-Hye ) - National Forensic Service Forensic DNA Division
 ( Park Hyun-Chul ) - National Forensic Service Forensic DNA Division
정주연 ( Jung Ju-Yeon ) - National Forensic Service Forensic DNA Division
진강남 ( Jin Gang-Nam ) - National Forensic Service Forensic DNA Division
황인관 ( Hwang In-Kwan ) - National Forensic Service Forensic DNA Division
강필원 ( Kang Pil-Won ) - National Forensic Service Forensic DNA Division

Abstract


Analysis techniques using DNA profiling are widely used in various fields including forensic science and new technologies such as the Direct PCR amplification method are being developed continuously in order to acquire the DNA profiles efficiently. However, it has a limits such as non-specific amplification according to the quality of crime scene evidence samples. Especially, split peaks caused by excessive DNA samples are one of the important factors that could cause the debate to allow researchers to interpret the DNA profile results.
In this study, we confirmed the occurrence rate of split peaks in each STR (short tandem repeats) locus of the GlobalFilerTM kit and investigated the possibility of improving the split peaks using several PCR additives such as DMSO (dimethylsulfoxide), MgCl2, Betaine and Tween-20. As a result, we could make three groups according to the occurrence rate of split peaks in Direct PCR and it was confirmed that the ratio of split peaks could be reduced by DMSO (87.4 %), MgCl2 (84.5 %) and Betaine (86.1 %), respectively. These results indicate that PCR additives such as DMSO, MgCl2 and Betaine can be improve the split peaks in Direct PCR and thereby facilitate subsequently a successful DNA profile results.

키워드

Direct PCR; PCR additive; split peaks; DNA profile; STR genotyping
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