AS-PCR

AS-PCR

Allele-specific polymerase chain reaction, see there.
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TNF-[alpha] -308G>A polymorphism was typed by AS-PCR as described previously (28,40).
The mutations in inhA-15 were determined by using 3 primers, namely TB92, TB93, and Rmut, via AS-PCR.
Several reports using AS-PCR achieved 100% detection rate in LPL but, curiously, some AS-PCR studies showed a detection rate below that achieved in our pyrosequencing assay.
Guerrero et al (69) developed an AS-PCR assay utilizing the T2134A mutation site in domain III S6 fragment of sodium channel gene.
Hence, we investigated the nucleotide polymorphism in the albendazole-binding domain of the isotype 1 [beta]-tubulin gene from several populations of Wuchereria bancrofti and developed an AS-PCR assay useful in screening for sensitive/resistance alleles among parasite populations and also evaluated its utility.
The general principle underlying the AS-PCR technique is to design a mutation-specific primer that produces the preferential amplification of a specific mutant allele (4).
To identify BRAF mutation, several analytic approaches are feasible including direct sequencing, (7) qPCR with melt curve analysis, (2,3) and AS-PCR.
Consenting participants were tested for CYP2D6 alleles by AS-PCR or PCR followed by restriction enzyme digestion at the University of Kentucky for the CYP2D6*3, *4, *5, *6, *7, *9, *17, *41, *1xn, *2xn/*35xn, and *4xn alleles based on published methods with some modifications (4, 9-16).
The nucleotides at positions 190, 481, 590, 803, and 857 were explored by digesting the 1211-bp PCR fragment or AS-PCR product carrying the wild-type or mutant allele with NciI, KpnI, TagI, DdeI, and BamHI, respectively.
Genotype Wild type Heterozygote Stop and Stop and Site transfer Direct transfer Direct 1 101/101 ND (a) 2/2 ND 2 201/201 201/201 18/18 18/18 3 88/88 88/88 7/7 7/7 4 94/94 ND 7/7 ND Total 484/484 289/289 34/34 25/25 Genotype Mutant All genotypes DNA Stop and Stop and extraction Site transfer Direct transfer Direct method 1 1/1 ND 104/104 ND Epicentre (b) or Qiagen (c) 2 1/1 1/1 220/220 220/220 Qiagen (c) 3 1/1 1/1 96/96 96/96 Qiagen (c) 4 1/1 ND 102/102 ND Bio-Rad (d) Total 4/4 2/2 522/522 316/316 Fluorescence Reference Site method method 1 Dynex Fluorolite PCR-RFLP 2 CytoFluor 4000 AS-PCR 3 CytoFluor 4000 PCR-RFLP 4 Dynex Fluorolite PCR-RFLP Total (a) ND, not determined.
A simple and inexpensive way to determine the genetic status of an individual is by the use of AS-PCR.