Nucleic Acids Research, Vol 26, Issue 7 1819-1825, Copyright © 1998 by Oxford University Press
MSB Judo, AB Wedel and C Wilson
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%.
ARTICLES
Stimulation and suppression of PCR-mediated recombination
Department of Biology and Center for the Molecular Biology of RNA, University of California, Santa Cruz, CA 95064, USA.
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