Detection of single DNA base differences by competitive oligonucleotide priming.
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.
Gibbs
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RA
Institute for Molecular Genetics, Baylor College of Medicine, Houston, TX 77030.
Nguyen
P N
PN
Caskey
C T
CT
D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D004247:DNA / Q000096:biosynthesis / Q000302:isolation & purification*; D004277:DNA, Single-Stranded; D004259:DNA-Directed DNA Polymerase; D005784:Gene Amplification; D051379:Mice; D008817:Mice, Mutant Strains; D008969:Molecular Sequence Data; D009154:Mutation; D009841:Oligonucleotides; D019914:Taq Polymerase; D013698:Templates, Genetic
D004277:DNA, Single-Stranded; D009841:Oligonucleotides; D004247:DNA; D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase