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Fluorescence Polarization in Homogeneous Nucleic Acid Analysis
Open accessAbstract
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 ∼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 http://www.genome.org.]
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Primer-Directed Enzymatic Amplification of DNA with a Thermostable DNA Polymerase
Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. J., Higuchi, R., Horn, G. T., Mullis, K. B., & Erlich, H. A. (1988) · Science · doi:10.1126/science.2448875 · Cited by 17,719
A novel MHC class I-like gene is mutated in patients with hereditary haemochromatosis
Feder, J. N., Gnirke, A., Thomas, W., Tsuchihashi, Z., Ruddy, D. A., Basava, A., Dormishian, F., Domingo, R., Ellis, M. C., Fullan, A., et al. (1996) · Nature Genetics · doi:10.1038/ng0896-399 · Cited by 3,664
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Vogelstein, B., & Kinzler, K. W. (1999) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.96.16.9236 · Cited by 1,770
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Kim, S., & Misra, A. (2007) · Annual Review of Biomedical Engineering · doi:10.1146/annurev.bioeng.9.060906.152037 · Cited by 622
High-Throughput Gene Mapping in Caenorhabditis elegans
Swan, K. A., Curtis, D. E., McKusick, K. B., Voinov, A. V., Mapa, F. A., & Cancilla, M. R. (2002) · Genome Research · doi:10.1101/gr.208902 · Cited by 616
Metadata
- Authors
- Chen, X., Levine, L., & Kwok, P.-Y.
- Venue
- Genome Research
- Publisher
- Cold Spring Harbor Laboratory
- Published
- 1999-05-01
- Volume
- 9
- Issue
- 5
- Pages
- 492-498
- ISSN
- 1088-9051 (ISSN-L)
- 1549-5469
- OpenAlex
- W2160437131
- Cited by
- 390
- References
- 2
- Type
- posted content
- Language
- en
- Open access
- bronze
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