Analysis of any point mutation in DNA. The amplification refractory mutation system (ARMS)

Abstract

We have improved the "polymerase chain reaction" (PCR) to permit rapid analysis of any known mutation in genomic DNA. We demonstrate a system, ARMS (Amplification Refractory Mutation System), that allows genotyping solely by inspection of reaction mixtures after agarose gel electrophoresis. The system is simple, reliable and non-isotopic. It will clearly distinguish heterozygotes at a locus from homozygotes for either allele. The system requires neither restriction enzyme digestion, allele-specific oligonucleotides as conventionally applied, nor the sequence analysis of PCR products. The basis of the invention is that unexpectedly, oligonucleotides with a mismatched 3'-residue will not function as primers in the PCR under appropriate conditions. We have analysed DNA from patients with alpha 1-antitrypsin (AAT) deficiency, from carriers of the disease and from normal individuals. Our findings are in complete agreement with allele assignments derived by direct sequencing of PCR products.

Keywords

BiologyGenotypingPolymerase chain reactionPoint mutationMolecular biologyOligonucleotidegenomic DNALocus (genetics)AlleleGeneticsDNALigase chain reactionMutationMultiplex polymerase chain reactionGenotypeGene

Affiliated Institutions

University of Wales GB

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Publication Info

Year: 1989
Type: article
Volume: 17
Issue: 7
Pages: 2503-2516
Citations: 2486
Access: Closed

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APA Style

                            
                                
                                    Charles R. Newton, 
                                
                                    Alexander Graham, 
                                
                                    Lesley Heptinstall
                                
                                et al.
                            
                            (1989). 
                            Analysis of any point mutation in DNA. The amplification refractory mutation system (ARMS). 
                            Nucleic Acids Research
                            , 17
                            (7)
                            , 2503-2516.
                            https://doi.org/10.1093/nar/17.7.2503
                        

Identifiers

DOI: 10.1093/nar/17.7.2503