allele-specific PCR

allele-specific PCR

The amplification of specific alleles, or DNA sequence variants, at the same locus by polymerase chain reaction. Specificity is achieved by designing one or both PCR primers so that they partially overlap the site of sequence difference between the amplified alleles.

AS-PCR is two-step nested PCR technique that allows detection of DNA with a point mutation in the presence of a 103–105-fold excess of normal DNA. AS-PCR is based on the observation that Taq polymerase will extend primers with a 3’ mismatch 103 to 106-fold less efficiently than perfectly matched primers; by choosing a primer that is complementary at its 3’ end to a point mutation, selective amplification of a mutant allele can be achieved.
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All the subjects were genotyped by allele-specific PCR. It was observed that the loci 6q25.1/rs6922269 and 9p21.3/rs1333049 were significantly associated with risk of CHD in this population (P=0.001 and 0.012, respectively).
The allele-specific PCR was performed using high throughput fluorescent based genotyping Kaspar assay.
Allele-specific PCR (AS-PCR) [2] is commonly used to discriminate between alleles that have single-nucleotide variants (SNVs) and well-defined insertions and deletions (1, 2).
Keywords: Parkinson's disease, a-Synuclein gene, Pakistani population, Allele-specific PCR, SNCA.
As MYD88 testing had now been added to the 2015 NCCN Guidelines as "useful in certain circumstances" and because he was at a research institution, allele-specific PCR of MYD88 was performed on the bone marrowbut was not detected.
Several recently developed methodologies, such as allele-specific amplification refractory mutation system PCR (ARMS), bead emulsification amplification and magnetics (BEAMing) technology, allele-specific PCR (AS-PCR), droplet digital PCR (ddPCR), and next generation sequencing (NGS), have been used to detect and quantify rare variants in the blood of melanoma patients, with analytical sensitivity ranging from 0.005 to 5% (Table 1).
The SNP was detected using allele-specific PCR. PCR master mix (# 12532-016, Invitrogen, Life Technologies, ThermoFisher Scientific, Waltham, MA, USA) was used for the SNP detection.
The Qiagen therascreen EGFR assay (14) utilizes a specific type of real-time PCR termed "allele-specific PCR".
Allele-specific PCR is a variant form of real-time PCR in which short oligonucleotide probes, specific for the wild-type and variant sequence, are mixed together in the amplification step.
KRAS mutations were identified in 12 pretreatment specimens, but only 9 posttreatment specimens, despite using laser capture microdissection to increase the proportion of tumor tissues and allele-specific PCR to improve the sensitivity of assays in the latter samples.