Citation: De Bin ZHU, Da XING, Xian LI, Lan ZHANG. A Novel Real-time Fluorescence Mutant-allele-specific Amplification Method for Rapid Single Nucleotide Polymorphism Analysis[J]. Chinese Chemical Letters, ;2006, 17(4): 499-501. shu

A Novel Real-time Fluorescence Mutant-allele-specific Amplification Method for Rapid Single Nucleotide Polymorphism Analysis

MOE Key Laboratory of Laser Life Science and Institute of Laser Life Science, South China Normal University, Guangzhou 510631

Received Date: 24 August 2005

Fund Project: This research is supported by the National Natural Science Foundation of China (60378043 30470494), and the Natural Science Foundation of Guangdong Province (015012 04010394).

Current methods for single nucleotide polymorphism (SNP) analysis are timeconsuming and complicated. We aimed at development of one-step real-time fluorescence mutant-allele-specific amplification (MASA) method for rapid SNP analysis. The method is a marriage of two technologies: MASA primers for target DNA and a double-stranded DNA-selective fluorescent dye, SYBR Green I. Genotypes are separated according to the different threshold cycles of the wild-type and mutant primers. K-ras oncogene was used as a target to validate the feasibility of the method. The experimental results showed that the different genotypes can be clearly discriminated by the assay. The real-time fluorescence MASA method will have an enormous potential for fast and reliable SNP analysis due to its simplicity and low cost.
- Mutant-allele-specific amplification,
- single nucleotide polymorphism analysis,
- SYBR Green I,
- K-ras oncogene
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