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Stimulation and suppression of PCR-mediated recombination
Open accessAbstract
Recombination, or chimera formation, is known to occur between related template sequences present in a single PCR amplification. To characterize the conditions under which such recombinant amplification products form we monitored the exchange of sequence between two homologous templates carrying different restriction sites separated by 282 bp. Using a typical cycling program the rates of recombination between the two restriction sites were 1 and 7% using Taq and Vent polymerases respectively over 12 doublings. However, by using long elongation times and cycling only to the mid-point of the amplification recombination could be suppressed below visual detection with both polymerases. Conversely, cycling programs designed to promote incomplete primer elongation and subsequent template strand exchange stimulated recombination to >20%.
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Cites
Primer-Directed Enzymatic Amplification of DNA with a Thermostable DNA Polymerase
Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. J., Higuchi, R., Horn, G. T., Mullis, K. B., & Erlich, H. A. (1988) · Science · doi:10.1126/science.2448875 · Cited by 17,719
Holland, J. H. (1992) · Scientific American · doi:10.1038/scientificamerican0792-66 · Cited by 5,764
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Alberdi, A., Aizpurua, O., Gilbert, M. T. P., & Bohmann, K. (2018) · Methods in Ecology and Evolution · doi:10.1111/2041-210x.12849 · Cited by 688
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Schnell, I. B., Bohmann, K., & Gilbert, M. T. P. (2015) · Molecular Ecology Resources · doi:10.1111/1755-0998.12402 · Cited by 630
Ban, Y., Honda, C., Hatsuyama, Y., Igarashi, M., Bessho, H., & Moriguchi, T. (2007) · Plant and Cell Physiology · doi:10.1093/pcp/pcm066 · Cited by 623
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Metadata
- Authors
- Judo, M. S. B., Wedel, A. B., & Wilson, C.
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 1998-04-01
- Volume
- 26
- Issue
- 7
- Pages
- 1819-1825
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- OpenAlex
- W2054662439
- Cited by
- 243
- References
- 2
- FWCI
- 2.36
- Type
- journal article
- Language
- en
- Open access
- bronze
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- Judo, M. S. B., Wedel, A. B., & Wilson, C. (1998). Stimulation and suppression of PCR-mediated recombination. Nucleic Acids Research. https://doi.org/10.1093/nar/26.7.1819
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
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