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.
Article
in Chinese
| 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>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Polymorphism, Restriction Fragment Length
/
Molecular Sequence Data
/
Base Sequence
/
Polymerase Chain Reaction
/
Reproducibility of Results
/
NAD(P)H Dehydrogenase (Quinone)
/
Polymorphism, Single Nucleotide
/
Genetics
/
Methods
Limits:
Adolescent
/
Adult
/
Female
/
Humans
/
Male
Language:
Chinese
Journal:
Chinese Journal of Medical Genetics
Year:
2008
Type:
Article
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