Enhanced discrimination of single nucleotide polymorphisms by artificial mismatch hybridization.
basic_science · Level V
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Abstract
In order to increase the discrimination of single nucleotide polymorphisms in DNA hybridization, artificial mismatches are inserted into probe oligonucleotides using the base analog 3-nitropyrrole. Differences in thermal stability (delta Tm) between hybrids formed with normal and single-nucleotide-variant DNA targets are increased by as much as 200% over conventional hybridization, and are strongly dependent upon the spacing between mismatches. The increased specificity is demonstrated by hybridization analysis and allele-specific amplification within the HLA-DRB locus.
Medical subject headings
- DNA
- Nucleic Acid Hybridization
- Polymorphism, Genetic