C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
150/8.5
C12Q 1/68 (2006.01) G01N 21/64 (2006.01) G01N 21/77 (2006.01) G01N 33/52 (2006.01)
Patent
CA 1299074
ABSTRACT OF THE INVENTION An assay for polynucleotides employing total inter- nally 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 pylynucleotides 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 curface through a spacer connected to an irre- versibly 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 poly- nucleotide 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. MJB-7
532664
Boehringer Mannheim Corporation
Macrae & Co.
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