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Applied and Environmental Microbiology, December 2004, p. 7545-7549, Vol. 70, No. 12
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.12.7545-7549.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

SHORT REPORT

Reevaluation and Reduction of a PCR Bias Caused by Reannealing of Templates

Shinya Kurata,1,2,3 Takahiro Kanagawa,2,3* Yukio Magariyama,4 Kyoko Takatsu,1,2 Kazutaka Yamada,1,2 Toyokazu Yokomaku,1 and Yoichi Kamagata2

Kankyo Engineering Co., Ltd., Kisarazu,1 Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba,2 Department of Molecular Engineering, Tokyo Institute of Technology, Nagatsuta, Midori-ku, Yokohama,3 Food Engineering Division, National Food Research Institute, Tsukuba, Ibaraki, Japan4

Received 26 December 2003/ Accepted 21 July 2004

ABSTRACT

We reevaluated the bias toward a 1:1 ratio of products in multitemplate PCR used in ecological studies and showed that the template reannealing at the annealing step would not cause the bias; however, the preferential homoduplex formation during temperature decrease from denaturation to annealing step would cause the bias.

* Corresponding author. Mailing address: National Institute of Advanced Industrial Science and Technology, Central 6, 1-1-1 Higashi, Tsukuba 305-8566, Japan. Phone: 81-29-861-6026. Fax: 81-29-861-6400. E-mail: kanagawa-taka{at}aist.go.jp.

Applied and Environmental Microbiology, December 2004, p. 7545-7549, Vol. 70, No. 12
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.12.7545-7549.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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