Patent ID: 6811973
Filing Date: 2004-11-02
Classification: C12Q

Abstract:
A method for quantifying the amount of a target molecule in solution comprising the steps of:a. incorporating one or more fluorescing nucleotide analogs into nucleotide probes to provide fluorescing nucleotide probes; b. providing a first substrate having a surface area; c. affixing a known number of said fluorescing nucleotide probes onto the substrate; d. detecting a first level of fluorescence from said fluorescing nucleotide probes on the substrate; e. contacting said first substrate with a sample solution comprising unlabeled target nucleotide sequences; f. providing sufficient conditions and time for unlabeled target molecules to selectively hybridize with fluorescing nucleotide probes on said substrate wherein hybridization of an unlabeled target molecule and an fluorescing nucleotide probe quenches fluorescence from said fluorescing nucleotide probe; g. removing the first substrate and detecting a second level of fluorescence from said fluorescing nucleotide probes after hybridization; h. repeating steps a. through g with subsequent substrates, having surface areas comprising known numbers of fluorescing nucleotide probes until all target molecules are hybridized and no longer quench said fluorescing nucleotide probes; and i. quantifying the amount of target molecule in the sample solution by adding the known number of fluorescing nucleotidc probes present on the first substrate and subsequent substrates contacted with and quenched by the unlabeled target molecule whereby the amount of the target molecule is quantified.