Direct haplotype determination by double ARMS: specificity, sensitivity and genetic applications.
We have developed a novel double Amplification Refractory Mutation System (double ARMS) using a highly polymorphic region 5' to the human delta-globin gene as a model system. The double ARMS approach involves using two allele-specific ARMS primers simultaneously during DNA amplification by the polymerase chain reaction (PCR). The resulting system is highly sensitive and more specific than single ARMS. In addition, this approach enables the elucidation of the relationship of polymorphic sites on the same chromosome and thus allows the direct determination of haplotypes. We have also demonstrated that this system can be used in conjunction with inverse PCR, the resulting double ARMS inverse PCR (DARMSI-PCR) may allow haplotype determination on polymorphic sites which are separated further apart than the length limit imposed by PCR. The double ARMS approach has numerous other applications in molecular biology including HLA typing, virology, forensic pathology and the investigation of the phenomenon of chimerism following bone marrow transplantation.
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Nuffield Department of Pathology and Bacteriology, John Radcliffe Hospital, Oxford, UK.
Patel
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Newton
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Markham
A F
AF
Fleming
K A
KA
Wainscoat
J S
JS
D000483:Alleles; D001483:Base Sequence; D015139:Blotting, Southern; D005914:Globins / Q000235:genetics; D006239:Haplotypes; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic / Q000235:genetics*; D015183:Restriction Mapping; D012680:Sensitivity and Specificity
D005914:Globins