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Nucleic Acids Symposium Series 2007 51(1):9-10; doi:10.1093/nass/nrm005
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© 2007 Oxford University Press

Development of an unnatural base pair for efficient PCR amplification

Ichiro Hirao1,*, Tsuneo Mitsui1, Michiko Kimoto1 and Shigeyuki Yokoyama1,2,3

1Protein Research Group, RIKEN Genomic Sciences Center, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan, 2Department of Biophysics and Biochemistry, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan, and 3RIKEN Harima Institute atSpring-8, 1-1-1 Kouto, Mikazuki-cho, Sayo, Hyogo 679-5148, Japan.

*Corresponding Author. ihirao{at}riken.jp

Abstract

An unnatural base pair system could expand the genetic alphabet, enabling the site-specific incorporation of extra, functional components into nucleic acids and proteins. We developed an unnatural base pair between 7-(2-thienyl)imidazo[4,5-b]pyridine (denoted by Ds) and 2-nitropyrrole (denoted by Pn), which specifically and efficiently functions in DNA amplification by PCR. After 20 cycles of PCR, the mutation rate of the Ds-Pn pair in an amplified DNA fragment was approximately 1%. The Ds-Pn pair system in combination with other unnatural base pairs could be useful for DNA/RNA-based biotechnology.


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