Modified Midi‐ and Mini‐Multiplex PCR Systems for Mitochondrial DNA Control Region Sequence Analysis in Degraded Samples |
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Authors: | Na Young Kim M.S. Hwan Young Lee Ph.D. Sun Joo Park Ph.D. Woo Ick Yang Ph.D. Kyoung‐Jin Shin Ph.D. |
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Affiliation: | 1. Department of Forensic Medicine, Yonsei University College of Medicine, , Seoul, 120‐752 Korea;2. Human Identification Research Center, Yonsei University, , Seoul, 120‐752 Korea;3. Department of Archaeology and Art History, Chungbuk National University College of Humanities, , Cheongju Chungbuk, 361‐763 Korea |
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Abstract: | Two multiplex polymerase chain reaction (PCR) systems (Midiplex and Miniplex) were developed for the amplification of the mitochondrial DNA (mtDNA) control region, and the efficiencies of the multiplexes for amplifying degraded DNA were validated using old skeletal remains. The Midiplex system consisted of two multiplex PCRs to amplify six overlapping amplicons ranging in length from 227 to 267 bp. The Miniplex system consisted of three multiplex PCRs to amplify 10 overlapping short amplicons ranging in length from 142 to 185 bp. Most mtDNA control region sequences of several 60‐year‐old and 400–500‐year‐old skeletal remains were successfully obtained using both PCR systems and consistent with those previously obtained by monoplex amplification. The multiplex system consisting of smaller amplicons is effective for mtDNA sequence analyses of ancient and forensic degraded samples, saving time, cost, and the amount of DNA sample consumed during analysis. |
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Keywords: | forensic science DNA typing mitochondrial DNA control region multiplex polymerase chain reaction small amplicon degraded DNA |
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