期刊
HUMAN MUTATION
卷 19, 期 4, 页码 315-323出版社
WILEY-LISS
DOI: 10.1002/humu.10058
关键词
SNP; mutation detection; genotyping; fluorescence polarization; nuclease assay; primer extension
资金
- NATIONAL EYE INSTITUTE [R01EY012557] Funding Source: NIH RePORTER
- NEI NIH HHS [R01-EY12557] Funding Source: Medline
When a fluorescent molecule is excited by plane polarized light, the fluorescence emitted is also polarized. The degree of fluorescence polarization (FP) detected, under constant temperature and solvent viscosity, is proportional to the molecular weight of the dye molecule. By monitoring the FP of a fluorescent dye, one can detect significant changes in the molecular weight of the molecule without separation or purification. Because the size of the probe is altered in the course of a number of single nucleotide polymorphism (SNP) genotyping reactions, FP is therefore an excellent detection mechanism for these assays. Indeed, FP detection can be used in SNP genotyping with the primer extension TaqMan(R) and Invader(R) assays. Use of FP detection makes it possible to reduce the cost of TaqMan(R) and Invader(R) probes by abrogating the need for a fluorescence quencher. Moreover, inexpensive, unpurified, and unlabeled probes are used in the primer extension reaction with FP detection. As an end-point detection mechanism, FP detection is suitable for high, throughput SNP genotyping. Hum Mutat 19:315-323, 2002. (C) 2002 Wiley-Liss, Inc.
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