DNA-based fluourescent sensor

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

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935 77, 935 78, C12Q 168, C12Q 100

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active

H00013986

ABSTRACT:
The fluorescent detection of target DNA is enhanced by amplifying the DNA in a sample by initiating a polymerase chain reaction using a first pair of primers and an appropriate polymerase. The DNA is then reamplified, this time using a second pair of nested primers which bracket DNA base sequences within the region bracketed by the first set of primers. At least one of the second pair of primers is bound to a fluorescent reporter molecule and at least one of the second pair of primers has bound thereto a specific binding site for a double-stranded DNA-binding protein. The thus amplified sample is then contacted with a fluorescent sensor having the appropriate double-stranded DNA-binding protein attached thereto. Then, the fluorescent reporter molecule is excited by light of the appropriate wavelength. The present invention provides greater sensitivity, adaptability and ease of use than previously available.

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Syvanen A. Medical Biology 64:313-324 (1986).
Meinkoth et al. Anal. Biochem. 138:267-84 (1984).
Colorimetric detection of specific DNA segments amplified by polymerase chain reactions, David J. Kemp et al., Proc. Natl. Acad. Sci. USA. vol. 86, pp. 2423-2427, Apr. 1989 Medical Sciences.
Fiber Optic-Based Immunosensors: A Progress Report, Suresh K. Bhatia et al., SPIE, vol. 1054, Fluorescence Detection III (1989), pp. 184-190.

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