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J Microbiol Methods. 2013 Jun;93(3):203-5. doi: 10.1016/j.mimet.2013.03.013. Epub 2013 Mar 26.

Slowing PCR ramp speed reduces chimera formation from environmental samples.

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  • 1Department of Biological Sciences, Campus Box 870344, University of Alabama, Tuscaloosa, AL 35487, United States.

Abstract

Chimeric sequences falsely increase the apparent diversity within samples. To examine chimera formation in PCR products from environmental DNA, clone libraries were prepared using different ramp speeds to reach the designated temperatures for each step of the PCR program. Slowing the thermocycler ramp speed to 1 °C s(-1) reduced chimera formation.

Copyright © 2013 Elsevier B.V. All rights reserved.

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