AS-PCR

AS-PCR

Allele-specific polymerase chain reaction, see there.
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Blood Group Genotyping PCR, PCR-RFLP, AS-PCR or PCR-SSP, Multiplex PCR, Real Time PCR, Sanger DNA Sequencing, Pyrosequencing Microarrays BeadChip Array, BloodChip, Genome Lab SNP Stream, Fluidic Microarray Systems, TaqMan OpenArray, MALDI-TOF-MS, Mini-Sequencing Blood Typing and Grouping Tests ABO, Antibody Panels, Antibody Screening/Indirect Antiglobulin, Antigen Typing (C, c, Duffy, E, e, I, i, Kell, Kidd, Le a, b, MN, P, S, s), Antiglobulin (Direct, C3 + IgG, IgG, C3), Crossmatching (Immediate Spin, Full Crossmatch), Rh (D, Du).
Guerrero et al (69) developed an AS-PCR assay utilizing the T2134A mutation site in domain III S6 fragment of sodium channel gene.
To identify BRAF mutation, several analytic approaches are feasible including direct sequencing, (7) qPCR with melt curve analysis, (2,3) and AS-PCR.
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 AS-PCR developed showed potential application as a tool for monitoring albendazole sensitivity/resistance alleles among W.
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).
We explored the effect of various conditions on AS-PCR amplification: for each AS-PCR, 3 mutation-specific oligonucleotides differing from each other by the position and type of mismatch were tested at the same concentration in a solution containing 2.
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.