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Ancient DNA as a Means to Investigate the European Neolithic
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Evolution, Genomics and Systematics, Evolutionary Biology.
2007 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The transition from a hunter-gatherer lifestyle to a farming lifestyle, i.e. the Neolithisation, is arguably the most important event in human prehistory. While the geography and dating of the Neolithisation is well known, the process is still under debate, especially if it occurred through diffusion of ideas or with migrating farmers. The process accelerated when alternative use of domesticated animals increased. Especially the use of dairy products, and the consumption of unprocessed milk, appears to be of importance. As milk consumption (lactose digestion) is dependent upon genetic components, it is debated whether the genetic disposition allowed for dairy production to evolve, or if the usages of dairy products added selection pressure that eventually lead to present day allele frequencies. Molecular genetics have the potential to solve this and similar questions, but only if the contamination problem, where authentic DNA can be distinguished from modern contaminating DNA, can be resolved.

Here I investigate the nature and extent of contamination with modern human DNA in museum specimens and explore several approaches to minimise this contamination and to authenticate DNA results from ancient humans. I use real-time quantification, pyrosequencing and FLX-generated clonal sequencing assays to generate data on ancient humans and ancient dogs. I further use the techniques to study the development of lactase persistence and the nature of animal domestication.

The results presented show that sample-based contamination is extensive, but can be minimised if treated with bleach. I retrieved authentic HVSI sequences from 30 Neolithic hunter-gatherers and farmers from Sweden, of which eighteen also yielded nuclear data indicating that the farmers had a higher frequency of the allele linked to lactase persistence compared to the hunter-gatherers. I conclude that genetic data from ancient humans as well as from ancient animals can be retrieved and used, but only under high stringency.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis , 2007. , p. 70
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 323
Keywords [en]
Molecular genetics, aDNA, contamination, authentication, mtDNA, Neolithisation
Keywords [sv]
Genetik
Identifiers
URN: urn:nbn:se:uu:diva-8162ISBN: 978-91-554-6940-5 (print)OAI: oai:DiVA.org:uu-8162DiVA, id: diva2:170526
Public defence
2007-09-21, Lindhalsalen, Norbyvägen 14, Uppsala, 13:00 (English)
Opponent
Supervisors
Available from: 2007-08-31 Created: 2007-08-31 Last updated: 2009-03-31Bibliographically approved
List of papers
1. Extensive human DNA contamination in extracts from ancient dog bones and teeth
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2005 (English)In: Molecular Biology and Evolution, ISSN 0737-4038, Vol. 22, no 10, p. 2040-2047Article in journal (Refereed) Published
Identifiers
urn:nbn:se:uu:diva-96064 (URN)
Available from: 2007-08-31 Created: 2007-08-31 Last updated: 2009-03-31Bibliographically approved
2. More on contamination: The use of asymmetric molecular behavior to identify authentic ancient human DNA
Open this publication in new window or tab >>More on contamination: The use of asymmetric molecular behavior to identify authentic ancient human DNA
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2007 (English)In: Molecular biology and evolution, ISSN 0737-4038, E-ISSN 1537-1719, Vol. 24, no 4, p. 998-1004Article in journal (Refereed) Published
Abstract [en]

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.

Keywords
contamination, ancient DNA, authentication
National Category
Biological Sciences
Identifiers
urn:nbn:se:uu:diva-96065 (URN)10.1093/molbev/msm015 (DOI)000245353200013 ()17255122 (PubMedID)
Available from: 2007-08-31 Created: 2007-08-31 Last updated: 2017-12-13Bibliographically approved
3. Barking up the wrong tree: Modern northern European dogs fail to explain their origin
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2008 (English)In: BMC Evolutionary Biology, ISSN 1471-2148, E-ISSN 1471-2148, Vol. 8, article id 71Article in journal (Refereed) Published
Abstract [en]

Background: Geographic distribution of the genetic diversity in domestic animals, particularly mitochondrial DNA, has often been used to infer centers of domestication. The underlying presumption is that phylogeographic patterns among domesticates were established during, or shortly after the domestication. Human activities are assumed not to have altered the haplogroup frequencies to any great extent. We studied this hypothesis by analyzing 24 mtDNA sequences in ancient Scandinavian dogs. Breeds originating in northern Europe are characterized by having a high frequency of mtDNA sequences belonging to a haplogroup rare in other populations (HgD). This has been suggested to indicate a possible origin of the haplogroup (perhaps even a separate domestication) in central or northern Europe. Results: The sequences observed in the ancient samples do not include the haplogroup indicative for northern European breeds (HgD). Instead, several of them correspond to haplogroups that are uncommon in the region today and that are supposed to have Asian origin. Conclusion: We find no evidence for local domestication. We conclude that interpretation of the processes responsible for current domestic haplogroup frequencies should be carried out with caution if based only on contemporary data. They do not only tell their own story, but also that of humans.

National Category
Biological Sciences
Identifiers
urn:nbn:se:uu:diva-96066 (URN)10.1186/1471-2148-8-71 (DOI)000254663500001 ()18307773 (PubMedID)
Available from: 2007-08-31 Created: 2007-08-31 Last updated: 2017-12-14Bibliographically approved
4. Ancient human DNA: authentication through FLX-sequenced PCR products in conjunction with bleach pre-treatment, quantitative PCR, assesment of assymetric behaviour and negative controls
Open this publication in new window or tab >>Ancient human DNA: authentication through FLX-sequenced PCR products in conjunction with bleach pre-treatment, quantitative PCR, assesment of assymetric behaviour and negative controls
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(English)Manuscript (Other (popular science, discussion, etc.))
Identifiers
urn:nbn:se:uu:diva-96067 (URN)
Available from: 2007-08-31 Created: 2007-08-31 Last updated: 2010-01-14Bibliographically approved
5. Different allele frequencies in the lactase gene in Scandinavian Neolithic populations and the development of dairy product consumtion
Open this publication in new window or tab >>Different allele frequencies in the lactase gene in Scandinavian Neolithic populations and the development of dairy product consumtion
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(English)Manuscript (Other (popular science, discussion, etc.))
Identifiers
urn:nbn:se:uu:diva-96068 (URN)
Available from: 2007-08-31 Created: 2007-08-31 Last updated: 2010-01-14Bibliographically approved

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