Skip Navigation

Nucleic Acids Research 2005 33(14):e126; doi:10.1093/nar/gni111
This Article
Right arrow Full Text Freely available
Right arrow Print PDF (1288K) Freely available
Right arrow Screen PDF (462K) Freely available
Right arrow Supplementary Material
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Shigemori, Y.
Right arrow Articles by Oishi, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Shigemori, Y.
Right arrow Articles by Oishi, M.
Related Collections
Right arrow Nucleic acid amplification
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Published online 8 August 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}oupjournals.org

Methods Online

Multiplex PCR: use of heat-stable Thermus thermophilus RecA protein to minimize non-specific PCR products

Yasushi Shigemori1,2, Tsutomu Mikawa3,4, Takehiko Shibata4 and Michio Oishi1,*

1Kazusa DNA Research Institute 2-6-7 Kazusa-kamatari, Kisarazu, Chiba 292-0818, Japan 2Aisin Cosmos R&D Co., Ltd 2-6-7 Kazusa-kamatari, Kisarazu, Chiba 292-0818, Japan 3RIKEN Harima Institute/SPring-8 Mikazuki cho, Hyogo 679-5148, Japan 4RIKEN Discovery Research Institute 2-1 Hirosawa, Wako, Saitama 351-0198, Japan

*To whom correspondence should be addressed. Tel: +81 438 52 3945; Fax: +81 438 52 3946; Email: oishi{at}kazusa.or.jp

Received May 28, 2005. Revised June 27, 2005. Accepted June 27, 2005.

In this paper we report that the inclusion of heat-resistant RecA protein from a thermophilic bacteria, Thermus thermophilus, and its cofactor (ATP) in PCR effectively eliminates non-specific PCR products. The effect of RecA protein, which catalyzes pairing between homologous DNA molecules with great fidelity in genetic recombination, is due to its promotion of precise priming in PCR (i.e. priming at sites where the primer sequence is completely complementary to that of the target sequence). In addition, the RecA protein substantially reduces the primer concentration required for PCR. These experimental results have led to the realization of multiplex PCR, which involves PCR for multiple sites in the same reaction mixture. We were able to successfully perform multiplex PCR with over a dozen reactions without affecting the amplification pattern of the PCR products.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Nucleic Acids ResHome page
A. V. Lebedev, N. Paul, J. Yee, V. A. Timoshchuk, J. Shum, K. Miyagi, J. Kellum, R. I. Hogrefe, and G. Zon
Hot Start PCR with heat-activatable primers: a novel approach for improved PCR performance
Nucleic Acids Res., September 16, 2008; (2008) gkn575v1.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
J. Inoue, M. Honda, S. Ikawa, T. Shibata, and T. Mikawa
The process of displacing the single-stranded DNA-binding protein from single-stranded DNA by RecO and RecR proteins
Nucleic Acids Res., January 17, 2008; 36(1): 94 - 109.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
F. Dahl, J. Stenberg, S. Fredriksson, K. Welch, M. Zhang, M. Nilsson, D. Bicknell, W. F. Bodmer, R. W. Davis, and H. Ji
Multigene amplification and massively parallel sequencing for cancer mutation discovery
PNAS, May 29, 2007; 104(22): 9387 - 9392.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
S. Fredriksson, J. Baner, F. Dahl, A. Chu, H. Ji, K. Welch, and R. W. Davis
Multiplex amplification of all coding sequences within 10 cancer genes by Gene-Collector
Nucleic Acids Res., April 1, 2007; 35(7): e47 - e47.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. Honda, J. Inoue, M. Yoshimasu, Y. Ito, T. Shibata, and T. Mikawa
Identification of the RecR Toprim Domain as the Binding Site for both RecF and RecO: A ROLE OF RecR IN RecFOR ASSEMBLY AT DOUBLE-STRANDED DNA-SINGLE-STRANDED DNA JUNCTIONS
J. Biol. Chem., July 7, 2006; 281(27): 18549 - 18559.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
K. K. Stanley and E. Szewczuk
Multiplexed tandem PCR: gene profiling from small amounts of RNA using SYBR Green detection
Nucleic Acids Res., November 24, 2005; 33(20): e180 - e180.
[Abstract] [Full Text] [PDF]


Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.