Korean J Phys Anthropol.
2007 Dec;20(4):267-281.
Development of Ancient DNA Isolation Method for Improved PCR Amplification
- Affiliations
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- 1Department of Science & Culture, Graduate School, Chung-Ang University, Korea. skull@cau.ac.kr
- 2Department of Microbiology, College of Medicine, Chung-Ang University, Korea.
- 3Department of Life science, College of Natural Science, Chung-Ang University, Korea.
- 4Department of Anthropology and Archeology, School of Social Sciences, National University of Mongolia, Ulaanbaatar, Mongolia.
- 5Department of Biology, Institute for Life Science, College of Natural Sciences, Kangwon National University, Korea.
- 6Department of Archaelogy and Art History, College of Humanities, Donga University, Korea.
- 7Division of Mathematical Science, College of Natural Sciences, Pukyong National University, Korea.
- 8DNA Analysis Section, National Institute of Scientific Investigation, Korea.
- 9Department of Laboratory Medicine, College of Medicine, Chung-Ang University, Korea.
- 10Department of Philosophy, College of Liberal Arts, Chung-Ang University, Korea.
- 11Department of Folklore Studies, Chung-Ang University, Korea.
- 12Department of Anatomy, College of Medicine, Chung-Ang University, Korea.
Abstract
- Ancient DNA analyses are widely used for evolutionary and phylogenetic study of mankind in anthropology and archeology. However, the DNA extraction from particularly poorly preserved ancient human samples is often unsuccessful in these analyses. In the present study, to improve the success rate of ancient DNA analysis, we introduced a high grade ancient DNA purification method using ion-exchange columns. We compared the success rate of ancient DNA analysis of this new method with that of the two methods that have been used for ancient DNA extraction, GENECLEAN(R) kit (Qbiogene) and Qiaquick column (Qiagen). Twelve ancient bone samples from Korea and Mongolia that are about 500 to 5,000 years old by an archeological estimation were used. As the DNA analysis methods, polymerase chain reaction (PCR) methods for the amplification of a mitochondrial DNA HV1 segment, a male sex determination marker DNA and M175 marker DNA that is used for the determination of O haplogroup of Y chromosome that is reportedly a common one in modern Korean people. The method developed in this study remarkably increased the success rate of DNA analysis compared with the other two methods. Using the GENECLEAN(R) kit, only two samples were amplifiable for the mitochondrial DNA, no samples for the male sex determination marker and M175 marker DNAs. Using the Qiaquick columns, nine samples were amplifiable for mitochondirial DNA, nine samples for male sex determination marker and six samples for M175 marker. The developed method allowed for the amplification of mitochondrial DNA from all samples, male sex determination marker from eight samples and M175 marker from eight samples. The results demonstrate that ion-exchange columns can be useful for the improved ancient DNA extraction in anthropology and archeology.
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