A homogeneous high-throughput genotyping method based on competitive hybridization.
Clin Biochem
; 36(8): 633-40, 2003 Nov.
Article
en En
| MEDLINE
| ID: mdl-14636879
ABSTRACT
OBJECTIVES:
A reliable high-throughput assay system is necessary for the analysis of the ever-increasing numbers of single-nucleotide polymorphisms (SNP) relevant to genetic screening studies. We describe an assay suitable also for large-scale screening programs. DESIGN ANDMETHODS:
The one-step assay is based on asymmetric PCR amplification of the target sequence and subsequent time-resolved fluorescence measurement. Asymmetric amplification results in a single-stranded PCR product that is detected in the amplification vessel with a highly sensitive, homogeneous hybridization method.RESULTS:
A dual label, homogeneous high-throughput platform for nucleic acid sequence analysis was developed and validated using a C/T single-nucleotide polymorphism in the insulin gene as a model analyte and applied also to two other SNP-assays (poliovirus receptor A/G-polymorphism and CD86-gene exon 2 A/G-polymorphism).CONCLUSIONS:
The described high-throughput genotyping technology is very competitive in price, simple in design and easily applied to any analyte sequence.
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Banco de datos:
MEDLINE
Asunto principal:
Polimorfismo de Nucleótido Simple
/
Hibridación de Ácido Nucleico
Tipo de estudio:
Prognostic_studies
Límite:
Humans
Idioma:
En
Revista:
Clin Biochem
Año:
2003
Tipo del documento:
Article
País de afiliación:
Finlandia