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Nucleic Acids Res. Apr 11, 1989; 17(7): 2437–2448.
PMCID: PMC317634
Detection of single DNA base differences by competitive oligonucleotide priming.
R A Gibbs, P N Nguyen, and C T Caskey
Institute for Molecular Genetics, Baylor College of Medicine, Houston, TX 77030.
Synthetic DNA oligonucleotides can serve as efficient primers for DNA synthesis even when there is a single base mismatch between the primers and the corresponding DNA template. However, when the primer-template annealing is carried out with a mixture of primers and at low stringency the binding of a perfectly matched primer is strongly favored relative to a primer differing by a single base. This primer competition is observed over a range of oligonucleotide sizes from twelve to sixteen bases and with a variety of base mismatches. When coupled with the polymerase chain reaction, for the amplification of specific DNA sequences, competitive oligonucleotide priming provides a simple general strategy for the detection of single DNA base differences.
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