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Nucleic Acids Research 2005 33(16):5219-5225; doi:10.1093/nar/gki840
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Published online 12 September 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oxfordjournals.org

Nucleic Acid Enzymes

High fidelity TNA synthesis by Therminator polymerase

Justin K. Ichida1,2,3, Allen Horhota4, Keyong Zou1,2,3, Larry W. McLaughlin4 and Jack W. Szostak1,2,3,*

1Howard Hughes Medical Institute, Massachusetts General Hospital Boston, MA 02114, USA 2Department of Molecular Biology, Massachusetts General Hospital Boston, MA 02114, USA 3Department of Genetics, Harvard Medical School Boston, MA 02115, USA 4Department of Chemistry, Boston College Chestnut Hill, MA 02467, USA

*To whom correspondence should be addressed. Tel: +1 617 726 5981; Fax: +1 617 726 6893; Email: szostak{at}molbio.mgh.harvard.edu

Received July 25, 2005. Revised August 30, 2005. Accepted August 30, 2005.

Therminator DNA polymerase is an efficient DNA-dependent TNA polymerase capable of polymerizing TNA oligomers of at least 80 nt in length. In order for Therminator to be useful for the in vitro selection of functional TNA sequences, its TNA synthesis fidelity must be high enough to preserve successful sequences. We used sequencing to examine the fidelity of Therminator-catalyzed TNA synthesis at different temperatures, incubation times, tNTP ratios and primer/template combinations. TNA synthesis by Therminator exhibits high fidelity under optimal conditions; the observed fidelity is sufficient to allow in vitro selection with TNA libraries of at least 200 nt in length.


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