Reduction of heteroduplex formation in PCR amplification |
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Authors: | E Michu M Mrá?ková B Vyskot and J ?lu?vová |
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Institution: | (1) Wisconsin National Primate Research Center, University of Wisconsin—Madison, Madison, WI 53706, USA;(2) Department of Pathology and Laboratory Medicine, University of Wisconsin—Madison, Madison, WI 53706, USA;(3) Battelle Biomedical Research Center, Columbus, OH 43201-2693, USA;(4) University of Wisconsin-Madison, 555 Science Drive, Madison, WI 53711, USA; |
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Abstract: | Heteroduplex formation is known to occur during mixed-template polymerase chain reaction (PCR) using universal primers, and
may represent a serious problem in several PCR-based analyses. A common way to eliminate heteroduplex formation is to use
reconditioning PCR. Because we detected that reconditioning PCR was not always sufficient to prevent heteroduplex formation,
we focused on developing methods for the elimination of heteroduplexes during PCR. We detected that the heteroduplex to homoduplex
ratio can be decreased by the addition of Taq polymerase and by a decrease in the number of PCR cycles. An appropriate combination of both of these approaches can be a
method generally applicable to decrease the formation of heteroduplexes. |
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