Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1992-01-02
1993-09-07
Chambers, Scott A.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
435 5, 436 63, 436 94, C12Q 168, C12Q 170, G01N 3300, G01N 3348
Patent
active
052427978
ABSTRACT:
An assay for polynucleotides employing total internal reflection of excitation radiation at a coating bonded to the surface of an optically conductive glass cell. The coating initially includes single-stranded polynucleotides coupled to individual attachment sites on the surface of the cell, such polynucleotides being complementary, at least in part to the single-stranded form of the polynucleotide that is being assayed. Each molecule of the coupled polynucleotide is connected to the cell surface through a spacer connected to an irreversibly conjugated polyadenine/polythymidine sequence at one end of the coupled polynucleotide.
When the surface of the coated cell is contacted with a sample that contains single-stranded polynucleotide complementary to the bound polynucleotide, renaturation will occur, forming a double-stranded form of the polynucleotide of interest. A fluorochrome dye specific to that double-stranded form is coupled to the latter, the dye including a chromophore that the excitation radiation will excite into fluorescence. The induced fluorescence is then gathered and measured.
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Dilica, K. Understanding DNA and Gene Cloning; John Wiley & Sons N.Y. 1984 p. 121.
Block Myron J.
Chambers Scott A.
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