Simultaneous Use of MutS and RecA for Suppression of Nonspecific Amplification during PCRReport as inadecuate




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Journal of Nucleic AcidsVolume 2013 2013, Article ID 823730, 5 pages

Research Article

RIKEN SPring-8 Center, Harima Institute, 1-1-1 Kouto, Sayo-cho, Sayo-gun, Hyogo 679-5148, Japan

Science and Technology Entrepreneurship Laboratory, Osaka University, Suita, Osaka 565-0871, Japan

Department of Biological Sciences, Graduate School of Science, Osaka University, 1-1 Machikaneyama-cho, Toyonaka, Osaka 560-0043, Japan

Received 27 March 2013; Accepted 21 June 2013

Academic Editor: Janusz M. Bujnicki

Copyright © 2013 Kenji Fukui and Seiki Kuramitsu. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Thermus thermophilus MutS, a thermostable mismatch-recognizing protein, is utilized in PCR to suppress nonspecific amplification by preventing synthesis from mismatched primers. T. thermophilus RecA also decreases nonspecific amplification by promoting proper hybridization between the primer and template. We observed that MutS and RecA function under the same reaction conditions and that MutS and RecA do not preclude each other. Furthermore, there were some DNA sequences for which only one of the 2 proteins effectively suppressed nonspecific amplification. The simultaneous use of MutS and RecA is a more attractive error-suppressing technique than the use of either of the 2 proteins alone.





Author: Kenji Fukui and Seiki Kuramitsu

Source: https://www.hindawi.com/



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