Dual Priming Oligonucleotide System for the Multiplex Detection of Respiratory Viruses and Snp Genotyping of Cyp2c19 Gene.
From: Seegene Institute of Life Science, 65-5 Bangyi-dong, Songpa-gu, Seoul 138-050, South Korea. chun@seegene.com
Nucleic acids research
- Publish Date: 2007
- ISSN: 1362-4962
- Volume: 35
- Issue: 6
- Pages: e40
- Medium: Internet
- Language: English
- Citation (JAMA): Chun Jong-Yoon, Kim Kyoung-Joong, Hwang In-Taek, et al. Dual Priming Oligonucleotide System for the Multiplex Detection of Respiratory Viruses and Snp Genotyping of Cyp2c19 Gene.. Nucleic Acids Res. 2007;35:e40
Abstract
Successful PCR starts with proper priming between an oligonucleotide primer and the template DNA. However, the inevitable risk of mismatched priming cannot be avoided in the currently used primer system, even though considerable time and effort are devoted to primer design and optimization of reaction conditions. Here, we report a novel dual priming oligonucleotide (DPO) which contains two separate priming regions joined by a polydeoxyinosine linker. The linker assumes a bubble-like structure which itself is not involved in priming, but rather delineates the boundary between the two parts of the primer. This structure results in two primer segments with distinct annealing properties: a longer 5’-segment that initiates stable priming, and a short 3’-segment that determines target-specific extension. This DPO-based system is a fundamental tool for blocking extension of non-specifically primed templates, and thereby generates consistently high PCR specificity even under less than optimal PCR conditions. The strength and utility of the DPO system are demonstrated here using multiplex PCR and SNP genotyping PCR.
Mesh Headings (Keywords): Animals, Aryl Hydrocarbon Hydroxylases, Coronavirus OC43, Human, DNA Primers, Genotype, Humans, Mice, Mixed Function Oxygenases, Orthomyxoviridae, Polymerase Chain Reaction, Polymorphism, Single Nucleotide, RNA Viruses, Respiratory Syncytial Virus, Human
Check for Full Text / PubMed Unique Identifier (PMID): 17287288
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