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SNP genotyping with fluorescence polarization detection
Author(s) -
Kwok PuiYan
Publication year - 2002
Publication title -
human mutation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 162
eISSN - 1098-1004
pISSN - 1059-7794
DOI - 10.1002/humu.10058
Subject(s) - snp genotyping , taqman , genotyping , fluorescence , biology , primer extension , fluorescence anisotropy , single nucleotide polymorphism , multiplex , molecular inversion probe , primer (cosmetics) , snp , microbiology and biotechnology , genetics , genotype , real time polymerase chain reaction , chemistry , dna , base sequence , gene , organic chemistry , quantum mechanics , membrane , physics
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 ® and Invader ® assays. Use of FP detection makes it possible to reduce the cost of TaqMan ® and Invader ® 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. © 2002 Wiley‐Liss, Inc.