Nucleic Acids Research, 1989, Vol. 17, No. 7 2503-2516
© 1989
MOLECULAR BIOLOGY |
Analysis of any point mutation in DNA. The amplification refractory mutation system (ARMS)
ICI Diagnostics, Gadbrook Park, Northwich, Cheshire CW9 7RA. ICI Cellmark Diagnostics, Blacklands Way, Abingdon Business Park, Abingdon, Oxon 0X14 IDY. 1University of Wales College of Medicine Department of Medical Biochemistry, Newport Road. Cardiff CF2 1SZ, UK and ICI Cellmark Diagnostics, 20271 Golden Rod Lane, Germantown, MD 20874, USA
Received February 23, 1989. Accepted March 2, 1989.
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, allelespecific 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 
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.
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