Mutant oligonucleotide extension amplification: a nonlabeling polymerase-chain-reaction-based assay for the detection of point mutations.
We have developed a new nonradioactive method for the identification of mutations causing beta-thalassemia and cystic fibrosis. The method is based on the priming of polymerase chain reaction (PCR) by an oligonucleotide complementary to the DNA sequence containing the mutation of interest, which anneals only to the perfectly matched sequence under high stringency conditions. A more upstream oligonucleotide primer is also included in the reaction to discriminate between homozygosity and heterozygosity for a particular mutation and serves as a control for the efficiency of the amplification.
Efremov
D G
DG
Research Center for New Technologies, Macedonian Academy of Sciences and Arts, Skopje, Yugoslavia.
Dimovski
A J
AJ
Janković
L
L
Efremov
G D
GD
D001483:Base Sequence; D003550:Cystic Fibrosis / Q000235:genetics*; D004247:DNA / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D009841:Oligonucleotides / Q000235:genetics*; D016133:Polymerase Chain Reaction; D013789:Thalassemia / Q000235:genetics*
D009841:Oligonucleotides; D004247:DNA