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      DNA recombination during PCR.

      Nucleic Acids Research
      Amino Acid Sequence, Base Sequence, Chimera, Cloning, Molecular, methods, DNA, Recombinant, metabolism, Gene Amplification, Gene Products, tat, genetics, Genes, Viral, Genes, tat, HIV-1, Molecular Sequence Data, Oligonucleotide Probes, Plasmids, Polymerase Chain Reaction, Recombination, Genetic, Sequence Homology, Nucleic Acid, tat Gene Products, Human Immunodeficiency Virus

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          Abstract

          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.

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