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Applied and Environmental Microbiology, August 2001, p. 3753-3755, Vol. 67, No. 8
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.8.3753-3755.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Optimization of Annealing Temperature To Reduce Bias Caused by a Primer Mismatch in Multitemplate PCR

Kousuke Ishii* and Manabu Fukui

Department of Biological Sciences, Graduate School of Science, Tokyo Metropolitan University, Hachiohji, Tokyo 192-0397, Japan

Received 7 February 2001/Accepted 14 May 2001

To reduce PCR bias derived from a primer mismatch, the effect of the annealing temperature on the product ratio was investigated by denaturing gradient gel electrophoresis analysis of PCR products from a mixture of perfect-match and one-mismatch templates. These templates were generated by PCR from Pediococcus acidilactici for one mismatch and Micrococcus luteus for the perfect match. PCRs showed that the bias was reduced at lower temperatures. An environmental sample was also examined.

* Corresponding author. Mailing address: Department of Biological Sciences, Graduate School of Science, Tokyo Metropolitan University, Hachiohji, Tokyo 192-0397, Japan. Phone: 81-426-77-2581. Fax: 81-426-77-2559. E-mail: ishii-kousuke{at}c.metro-u.ac.jp.

Applied and Environmental Microbiology, August 2001, p. 3753-3755, Vol. 67, No. 8
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.8.3753-3755.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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