Enhanced discrimination of single nucleotide polymorphisms by artificial mismatch hybridization.
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.
Guo
Z
Z
Department of Chemistry, University of Wisconsin-Madison 53706, USA.
Liu
Q
Q
Smith
L M
LM
D000483:Alleles; D001483:Base Sequence; D001709:Biotechnology; D004247:DNA / Q000737:chemistry* / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D004355:Drug Stability; D006684:HLA-DR Antigens / Q000235:genetics; D006801:Humans; D066298:In Vitro Techniques; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes / Q000737:chemistry / Q000235:genetics; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D011758:Pyrroles / Q000737:chemistry; D013816:Thermodynamics
D017931:DNA Primers; D006684:HLA-DR Antigens; D015345:Oligonucleotide Probes; D011758:Pyrroles; C094911:3-nitropyrrole; D004247:DNA