37
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      Post Mortem DNA Degradation of Human Tissue Experimentally Mummified in Salt

      research-article

      Read this article at

      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          Mummified human tissues are of great interest in forensics and biomolecular archaeology. The aim of this study was to analyse post mortem DNA alterations in soft tissues in order to improve our knowledge of the patterns of DNA degradation that occur during salt mummification. In this study, the lower limb of a female human donor was amputated within 24 h post mortem and mummified using a process designed to simulate the salt dehydration phase of natural or artificial mummification. Skin and skeletal muscle were sampled at multiple time points over a period of 322 days and subjected to genetic analysis. Patterns of genomic fragmentation, miscoding lesions, and overall DNA degradation in both nuclear and mitochondrial DNA was assessed by different methods: gel electrophoresis, multiplex comparative autosomal STR length amplification, cloning and sequence analysis, and PCR amplification of different fragment sizes using a damage sensitive recombinant polymerase. The study outcome reveals a very good level of DNA preservation in salt mummified tissues over the course of the experiment, with an overall slower rate of DNA fragmentation in skin compared to muscle.

          Related collections

          Most cited references18

          • Record: found
          • Abstract: found
          • Article: not found

          Detection and quantification of rare mutations with massively parallel sequencing.

          The identification of mutations that are present in a small fraction of DNA templates is essential for progress in several areas of biomedical research. Although massively parallel sequencing instruments are in principle well suited to this task, the error rates in such instruments are generally too high to allow confident identification of rare variants. We here describe an approach that can substantially increase the sensitivity of massively parallel sequencing instruments for this purpose. The keys to this approach, called the Safe-Sequencing System ("Safe-SeqS"), are (i) assignment of a unique identifier (UID) to each template molecule, (ii) amplification of each uniquely tagged template molecule to create UID families, and (iii) redundant sequencing of the amplification products. PCR fragments with the same UID are considered mutant ("supermutants") only if ≥95% of them contain the identical mutation. We illustrate the utility of this approach for determining the fidelity of a polymerase, the accuracy of oligonucleotides synthesized in vitro, and the prevalence of mutations in the nuclear and mitochondrial genomes of normal cells.
            Bookmark
            • Record: found
            • Abstract: not found
            • Article: not found

            Ancient DNA: do it right or not at all.

              Bookmark
              • Record: found
              • Abstract: found
              • Article: not found

              A novel strategy to engineer DNA polymerases for enhanced processivity and improved performance in vitro.

              Mechanisms that allow replicative DNA polymerases to attain high processivity are often specific to a given polymerase and cannot be generalized to others. Here we report a protein engineering-based approach to significantly improve the processivity of DNA polymerases by covalently linking the polymerase domain to a sequence non-specific dsDNA binding protein. Using Sso7d from Sulfolobus solfataricus as the DNA binding protein, we demonstrate that the processivity of both family A and family B polymerases can be significantly enhanced. By introducing point mutations in Sso7d, we show that the dsDNA binding property of Sso7d is essential for the enhancement. We present evidence supporting two novel conclusions. First, the fusion of a heterologous dsDNA binding protein to a polymerase can increase processivity without compromising catalytic activity and enzyme stability. Second, polymerase processivity is limiting for the efficiency of PCR, such that the fusion enzymes exhibit profound advantages over unmodified enzymes in PCR applications. This technology has the potential to broadly improve the performance of nucleic acid modifying enzymes.
                Bookmark

                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2014
                22 October 2014
                : 9
                : 10
                : e110753
                Affiliations
                [1 ]Institute of Evolutionary Medicine, University of Zurich, Zurich, Switzerland
                [2]Department of Forensic Genetics, Institute of Legal Medicine, University of Zürich, Zürich, Switzerland
                [3 ]Division of Cell- and Neurobiology, Institute of Anatomy, University of Zurich, Zurich, Switzerland
                Kliniken der Stadt Köln gGmbH, Germany
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: NS CP FR. Performed the experiments: GA KP NS CH. Analyzed the data: NS CW AB CH. Contributed reagents/materials/analysis tools: FR CH. Wrote the paper: NS CW AB.

                [¤a]

                Current address: Department of History and Ethnology, Demokritus University of Thrace, Komotini, Greece

                [¤b]

                Current address: Molecular Anthropology Laboratories, Department of Anthropology, University of Oklahoma, Norman, Oklahoma, United States of America

                Article
                PONE-D-14-20088
                10.1371/journal.pone.0110753
                4206501
                25337822
                5bc3fa19-6177-4680-b1ae-ee07ce0157ca
                Copyright @ 2014

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 27 June 2014
                : 22 September 2014
                Page count
                Pages: 10
                Funding
                Funding provided by Swiss National Science Foundation (120662) http://www.snf.ch/de/Seiten/default.aspx, Mäxi Stiftung (FR). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and life sciences
                Biochemistry
                DNA
                Ancient DNA
                Genetics
                Human Genetics
                Mitochondrial Genetics
                Molecular Genetics
                Mutation
                Molecular Biology
                Paleontology
                Paleogenetics
                Physical Anthropology
                Custom metadata
                The authors confirm that all data underlying the findings are fully available without restriction. Ethical and legal restrictions make our data unsuitable for public deposition. An anonymized raw-data set is available to interested researchers upon request to Prof. Frank Rühli ( frank.ruehli@ 123456iem.uzh.ch ).

                Uncategorized
                Uncategorized

                Comments

                Comment on this article