Molecular breeding of polymerases for amplification of ancient DNA View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2007-08

AUTHORS

Marc d'Abbadie, Michael Hofreiter, Alexandra Vaisman, David Loakes, Didier Gasparutto, Jean Cadet, Roger Woodgate, Svante Pääbo, Philipp Holliger

ABSTRACT

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. More... »

PAGES

939-943

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/nbt1321

DOI

http://dx.doi.org/10.1038/nbt1321

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1047090781

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/17632524


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