Establishing a single-tube fluorescent bidirectional PCR method to detect the 609C/T polymorphism of NQO1 gene / 中华医学遗传学杂志
Chinese Journal of Medical Genetics
;
(6): 230-232, 2008.
Artigo
em Chinês
| WPRIM
| ID: wpr-229783
ABSTRACT
<p><b>OBJECTIVE</b>To develop a single-tube fluorescent bidirectional PCR method to detect the 609C/T polymorphism of NAD(P)H quinone oxidoreductase 1 (NQO1) gene.</p><p><b>METHODS</b>Two primers of NQO1 gene C609T locus were designed. Using these primers, a SYBR Green I fluorescent bidirectional PCR, combined with melting curve analysis of the PCR products, were optimized to differentiate the 609C/T polymorphisms in 191 samples of human genomic DNA. The accuracy of the fluorescent bidirectional PCR was validated by the classical method of PCR-restriction fragment length polymorphism(RFLP) in 62 of these 191 samples.</p><p><b>RESULTS</b>In the 62 samples, the genotypes determined by the fluorescent bidirectional PCR were 100% consistent with the ones by the PCR-RFLP. The frequencies of genotypes of homozygous wild-type (CC), heterozygous (CT), and homozygous mutant (TT) were 28%, 50%, and 22%, respectively, in the 191 samples.</p><p><b>CONCLUSION</b>The single-tube fluorescent bidirectional PCR method established here provides a simple, rapid, accurate and inexpensive assay to determine the 609C/T polymorphism of NQO1 gene. The assay is suitable to detect the single nucleotide polymorphism in large-scale samples.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Polimorfismo de Fragmento de Restrição
/
Dados de Sequência Molecular
/
Sequência de Bases
/
Reação em Cadeia da Polimerase
/
Reprodutibilidade dos Testes
/
NAD(P)H Desidrogenase (Quinona)
/
Polimorfismo de Nucleotídeo Único
/
Genética
/
Métodos
Limite:
Adolescente
/
Adulto
/
Feminino
/
Humanos
/
Masculino
Idioma:
Chinês
Revista:
Chinese Journal of Medical Genetics
Ano de publicação:
2008
Tipo de documento:
Artigo
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