| Molecular breeding of polymerases for amplification of ancient DNA. | |
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MedLine Citation:
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PMID: 17632524 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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In the absence of repair, lesions accumulate in DNA. Thus, DNA persisting in specimens of paleontological, archaeological or forensic interest is inevitably damaged. We describe a strategy for the recovery of genetic information from damaged DNA. By molecular breeding of polymerase genes from the genus Thermus (Taq (Thermus aquaticus), Tth (Thermus thermophilus) and Tfl (Thermus flavus)) and compartmentalized self-replication selection, we have evolved polymerases that can extend single, double and even quadruple mismatches, process non-canonical primer-template duplexes and bypass lesions found in ancient DNA, such as hydantoins and abasic sites. Applied to the PCR amplification of 47,000-60,000-year-old cave bear DNA, these outperformed Taq DNA polymerase by up to 150% and yielded amplification products at sample dilutions at which Taq did not. Our results demonstrate that engineered polymerases can expand the recovery of genetic information from Pleistocene specimens and may benefit genetic analysis in paleontology, archeology and forensic medicine. |
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Authors:
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Marc d'Abbadie; Michael Hofreiter; Alexandra Vaisman; David Loakes; Didier Gasparutto; Jean Cadet; Roger Woodgate; Svante Pääbo; Philipp Holliger |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Intramural; Research Support, Non-U.S. Gov't Date: 2007-07-15 |
Journal Detail:
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Title: Nature biotechnology Volume: 25 ISSN: 1087-0156 ISO Abbreviation: Nat. Biotechnol. Publication Date: 2007 Aug |
Date Detail:
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Created Date: 2007-08-09 Completed Date: 2007-11-13 Revised Date: 2009-11-18 |
Medline Journal Info:
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Nlm Unique ID: 9604648 Medline TA: Nat Biotechnol Country: United States |
Other Details:
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Languages: eng Pagination: 939-43 Citation Subset: IM |
Affiliation:
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Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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DNA
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genetics* DNA Damage / genetics* DNA Fingerprinting / methods* DNA-Directed DNA Polymerase / chemistry*, genetics Forensic Genetics / methods Fossils Polymerase Chain Reaction / methods* Sequence Analysis, DNA / methods* Specimen Handling / methods* |
| Grant Support | |
ID/Acronym/Agency:
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U.1051.03.006(78804)//Medical Research Council |
| Chemical | |
Reg. No./Substance:
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9007-49-2/DNA; EC 2.7.7.7/DNA-Directed DNA Polymerase |
| Comments/Corrections | |
Comment In:
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Nat Biotechnol. 2007 Aug;25(8):872-4
[PMID:
17687362
]
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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