DNA recombination during PCR

PCR co-amplification of two distinct HIV1 tat gene sequences lead to the formation of recombinant DNA molecules. The frequency of such recombinants, up to 5.4% of all amplified molecules, could be decreased 2.7 fold by a 6 fold increase in Taq DNA polymerase elongation time. Crossover sites mapped e...

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Veröffentlicht in:Nucleic acids research 1990-04, Vol.18 (7), p.1687-1691
Hauptverfasser: MEYERHANS, A, VARTANIAN, J.-P, WAIN-HOBSON, S
Format: Artikel
Sprache:eng
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Zusammenfassung:PCR co-amplification of two distinct HIV1 tat gene sequences lead to the formation of recombinant DNA molecules. The frequency of such recombinants, up to 5.4% of all amplified molecules, could be decreased 2.7 fold by a 6 fold increase in Taq DNA polymerase elongation time. Crossover sites mapped essentially to three discrete regions suggesting specific Taq DNA polymerase pause or termination sites. PCR mediated recombination may be a problem when studying heterogeneous genetic material such as RNA viruses, multigene families, or repetitive sequences. This phenomenon can be exploited to create chimeric molecules from related sequences.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/18.7.1687