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Allele-specific PCR (AS-PCR) (CAT#: STEM-MB-0198-WXH)

Introduction

Allele-specific polymerase chain reaction (AS-PCR) is a technique based on allele-specific primers, which can be used to analyze single nucleotide polymorphism(SNP). A single nucleotide polymorphism (abbreviated SNP, pronounced snip) is a genomic variant at a single base position in the DNA. The allele-specific PCR is also called the (amplification refractory mutation system) ARMS-PCR.




Principle

Allele-specific PCR uses two different primers for two different alleles. One is the mutant set of primers which are refractory to the normal PCR, and the other is the normal set of primers, which are refractory to the mutant PCR reaction. The 3’ ends of these primers are modified such that one set of the primer can amplify the normal allele while others amplify the mutant allele. This mismatch allows the primer to amplify a single allele.

Applications

• SNP Genotyping.
• Detection of the single gene point mutation such as sickle cell anemia and thalassemia.
• The direct determination of ABO blood group genotypes.
• DNA-based diagnostic involving the diagnosis of genetic and infectious diseases.

Procedure

1.Primer design
2.PCR amplification
3.Gel electrophoresis