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MBE Advance Access originally published online on January 25, 2007
Molecular Biology and Evolution 2007 24(4):998-1004; doi:10.1093/molbev/msm015
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© The Author 2007. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

Research Articles

More on Contamination: The Use of Asymmetric Molecular Behavior to Identify Authentic Ancient Human DNA

Helena Malmström*, Emma M. Svensson*, M. Thomas P. Gilbert{dagger}, Eske Willerslev{dagger}, Anders Götherström* and Gunilla Holmlund{ddagger}

* Evolutionary Biology, Uppsala University, Uppsala, Sweden
{dagger} Centre for Ancient Genetics, Niels Bohr Institute and Biological Institute, University of Copenhagen, Copenhagen, Denmark
{ddagger} The National Board of Forensic Medicine, Department of Forensic Genetics and Forensic Toxicology, Linköping, Sweden

E-mail: anders.gotherstrom{at}ebc.uu.se.

Accepted for publication January 22, 2007.

Authentication of ancient human DNA results is an exceedingly difficult challenge due to the presence of modern contaminant DNA sequences. Nevertheless, the field of ancient human genetics generates huge scientific and public interest, and thus researchers are rarely discouraged by problems concerning the authenticity of such data. Although several methods have been developed to the purpose of authenticating ancient DNA (aDNA) results, while they are useful in faunal research, most of the methods have proven complicated to apply to ancient human DNA. Here, we investigate in detail the reliability of one of the proposed criteria, that of appropriate molecular behavior. Using real-time polymerase chain reaction (PCR) and pyrosequencing, we have quantified the relative levels of authentic aDNA and contaminant human DNA sequences recovered from archaeological dog and cattle remains. In doing so, we also produce data that describes the efficiency of bleach incubation of bone powder and its relative detrimental effects on contaminant and authentic ancient DNA. We note that bleach treatment is significantly more detrimental to contaminant than to authentic aDNA in the bleached bone powder. Furthermore, we find that there is a substantial increase in the relative proportions of authentic DNA to contaminant DNA as the PCR target fragment size is decreased. We therefore conclude that the degradation pattern in aDNA provides a quantifiable difference between authentic aDNA and modern contamination. This asymmetrical behavior of authentic and contaminant DNA can be used to identify authentic haplotypes in human aDNA studies.

Key Words: contamination • ancient DNA • authentication


Lisa Matisoo-Smith, Associate Editor


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