Статья

Patterns of nucleotide misincorporations during enzymatic amplification and direct large-scale sequencing of ancient DNA

M. Stiller, R. Green, M. Ronan, J. Simons, L. Du, W. He, M. Egholm, J. Rothberg, S. Keats, N. Ovodov, E. Antipina, G. Baryshnikov, Y. Kuzmin, A. Vasilevski, G. Wuenschell, J. Termini, M. Hofreiter, V. Jaenicke-Després, S. Pääbo,
2021

Whereas evolutionary inferences derived from present-day DNA sequences are by necessity indirect, ancient DNA sequences provide a direct view of past genetic variants. However, base lesions that accumulate in DNA over time may cause nucleotide misincorporations when ancient DNA sequences are replicated. By repeated amplifications of mitochondrial DNA sequences from a large number of ancient wolf remains, we show that C/G-to-T/A transitions are the predominant type of such misincorporations. Using a massively parallel sequencing method that allows large numbers of single DNA strands to be sequenced, we show that modifications of C, as well as to a lesser extent of G, residues cause such misincorporations. Experiments where oligonucleotides containing modified bases are used as templates in amplification reactions suggest that both of these types of misincorporations can be caused by deamination of the template bases. New DNA sequencing methods in conjunction with knowledge of misincorporation processes have now, in principle, opened the way for the determination of complete genomes from organisms that became extinct during and after the last glaciation. © 2006 by The National Academy of Sciences of the USA.

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  • 1. Version of Record от 2021-04-27

Метаданные

Об авторах
  • M. Stiller
    Max Planck Institute for Evolutionary Anthropology, D-04103 Leipzig, Germany
  • R. Green
    454 Life Sciences, Branford, CT 06405, United States
  • M. Ronan
    P.O. Box 350, London WC1, United Kingdom
  • J. Simons
    Institute of Archaeology and Ethnography, Siberian Branch, Russian Academy of Sciences, Krasnoyarsk 660049, Russian Federation
  • L. Du
    Institute of Archaeology, Russian Academy of Sciences, Moscow 117036, Russian Federation
  • W. He
    Zoological Institute, Russian Academy of Sciences, St. Petersburg 199034, Russian Federation
  • M. Egholm
    Pacific Institute of Geography, Far Eastern Branch, Russian Academy of Sciences, Vladivostok 690041, Russian Federation
  • J. Rothberg
    Sakhalin State University, Yuzhno-Sakhalinsk 693008, Russian Federation
  • S. Keats
    Division of Molecular Biology, Beckman Research Institute of the City of Hope, Duarte, CA 91010, United States
  • N. Ovodov
  • E. Antipina
  • G. Baryshnikov
  • Y. Kuzmin
  • A. Vasilevski
  • G. Wuenschell
  • J. Termini
  • M. Hofreiter
  • V. Jaenicke-Després
  • S. Pääbo
Название журнала
  • Proceedings of the National Academy of Sciences of the United States of America
Том
  • 103
Выпуск
  • 37
Страницы
  • 13578-13584
Ключевые слова
  • DNA; DNA base; mitochondrial DNA; article; deamination; DNA damage; DNA extraction; DNA sequence; DNA strand; gene amplification; nonhuman; nucleotide sequence; priority journal; wolf; Canidae
Издатель
  • National Academy of Sciences
Тип документа
  • journal article
Источник
  • scopus