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Int. J. Mol. Sci. 2013, 14(3), 6436-6453; doi:10.3390/ijms14036436

Thermostable Mismatch-Recognizing Protein MutS Suppresses Nonspecific Amplification during Polymerase Chain Reaction (PCR)

1
RIKEN SPring-8 Center, Harima Institute, 1-1-1 Kouto, Sayo-cho, Sayo-gun, Hyogo 679-5148, Japan
2
Department of Biological Sciences, Graduate School of Science, Osaka University, 1-1, Machikaneyama-cho, Toyonaka, Osaka 560-0043, Japan
3
RIKEN Systems and Structural Biology Center, 1-7-22 Suehiro-cho, Tsurumi, Yokohama 230-0045, Japan
*
Author to whom correspondence should be addressed.
Received: 31 January 2013 / Revised: 28 February 2013 / Accepted: 11 March 2013 / Published: 21 March 2013
(This article belongs to the Special Issue Molecular Cut and Paste)
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Abstract

Polymerase chain reaction (PCR)-related technologies are hampered mainly by two types of error: nonspecific amplification and DNA polymerase-generated mutations. Here, we report that both errors can be suppressed by the addition of a DNA mismatch-recognizing protein, MutS, from a thermophilic bacterium. Although it had been expected that MutS has a potential to suppress polymerase-generated mutations, we unexpectedly found that it also reduced nonspecific amplification. On the basis of this finding, we propose that MutS binds a mismatched primer-template complex, thereby preventing the approach of DNA polymerase to the 3' end of the primer. Our simple methodology improves the efficiency and accuracy of DNA amplification and should therefore benefit various PCR-based applications, ranging from basic biological research to applied medical science.
Keywords: polymerase chain reaction; DNA mismatch repair; mismatch-recognizing protein; Thermus thermophilus; Aquifex aeolicus polymerase chain reaction; DNA mismatch repair; mismatch-recognizing protein; Thermus thermophilus; Aquifex aeolicus
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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MDPI and ACS Style

Fukui, K.; Bessho, Y.; Shimada, A.; Yokoyama, S.; Kuramitsu, S. Thermostable Mismatch-Recognizing Protein MutS Suppresses Nonspecific Amplification during Polymerase Chain Reaction (PCR). Int. J. Mol. Sci. 2013, 14, 6436-6453.

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