Journal
BIOLOGIA PLANTARUM
Volume 54, Issue 1, Pages 173-176Publisher
SPRINGER
DOI: 10.1007/s10535-010-0029-8
Keywords
polymerase chain reaction; reconditioning PCR; mixed-template PCR
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Funding
- Institutional Research Plans [AVOZ50040507, AV0Z50040702]
- Grant Agency of the Czech Republic
- Grant Agency of AS CR [KJB600040801]
- Czech Ministry of Education [LC06004]
- [204/09/H002]
- [IAA600040801]
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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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