Fluorescence Polarization in Homogeneous Nucleic Acid Analysis | Zendy

Xiangning Chen | Zendy; Leanna Levine | Zendy; PuiYan Kwok | Zendy

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Fluorescence Polarization in Homogeneous Nucleic Acid Analysis

Author(s) -

Xiangning Chen,

Leanna Levine,

PuiYan Kwok

Publication year - 1999

Publication title -

genome research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.556

H-Index - 297

eISSN - 1549-5469

pISSN - 1088-9051

DOI - 10.1101/gr.9.5.492

Subject(s) - biology , microbiology and biotechnology , primer extension , dna , oligonucleotide , taq polymerase , genotyping , primer dimer , primer (cosmetics) , fluorophore , nucleic acid , multiplex ligation dependent probe amplification , sequencing by ligation , genomic dna , fluorescence , polymerase chain reaction , multiplex polymerase chain reaction , genetics , dna polymerase , genomic library , gene , genotype , chemistry , base sequence , thermus aquaticus , physics , organic chemistry , quantum mechanics , exon

A new method for DNA diagnostics based on template-directed primer extension and detection by fluorescence polarization is described. In this method, amplified genomic DNA containing a polymorphic locus is incubated with oligonucleotide primers (designed to hybridize to the DNA template adjacent to the polymorphic site) in the presence of allele-specific dye-labeled dideoxyribonucleoside triphosphates and a commercially available modified Taq DNA polymerase. The primer is extended by the dye-terminator specific for the allele present on the template, increasing approximately 10-fold the molecular weight of the fluorophore. At the end of the reaction, the fluorescence polarization of the two dye-terminators in the reaction mixture are analyzed directly without separation or purification. This homogeneous DNA diagnostic method is shown to be highly sensitive and specific and is suitable for automated genotyping of large number of samples. [The data shown in Figure 3 are available as an online supplement at.]

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