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Multiplex fluorescence-based primer extension method for quantitative mutation analysis of mitochondrial DNA and its diagnostic application for Alzheimer's disease

被引：30
作者：
Fahy, E
Nazarbaghi, R
Zomorrodi, M
Herrnstadt, C
Parker, WD
Davis, RE
Ghosh, SS
机构：
[1] MITOKOR,SAN DIEGO,CA 92121
[2] UNIV VIRGINIA,SCH MED,DEPT NEUROL,CHARLOTTESVILLE,VA 22908
[3] UNIV VIRGINIA,SCH MED,DEPT PEDIAT,CHARLOTTESVILLE,VA 22908
来源：
NUCLEIC ACIDS RESEARCH | 1997年 / 25卷 / 15期
关键词：
D O I：
10.1093/nar/25.15.3102
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
A sensitive and highly reproducible multiplexed primer extension assay is described for quantitative mutation analysis of heterogeneous DNA populations. Wild-type and mutant target DNA are simultaneously probed in competitive primer extension reactions using fluorophor-labeled primers and high fidelity thermostable DNA polymerases in the presence of defined mixtures of deoxy- and dideoxynucleotides, Primers are differentially extended and the resulting products are distinguished by size and dye label. Wild-type:mutant DNA ratios are determined from the fluorescence intensities associated with electrophoretically resolved reaction products, Multiple nucleotide sites can be simultaneously interrogated with uniquely labeled primers of different lengths, The application of this quantitative technique is shown in the analysis of heteroplasmic point mutations in mitochondrial DNA that are associated with Alzheimer's disease.
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收藏
页码：3102 / 3109
页数：8
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