Appeal No. 1999-2814
Application No. 08/990,539
(b) adding said first probe to a sample that contains said first
fragment to form a first mixture containing a first reaction
product of said first probe and said first fragment;
(c) exposing said first mixture to an exciting amount of radiation;
(d) detecting fluorescence of said first metal-ligand complex;
(e) identifying a first base sequence of said first fragment based on
fluorescence of said first metal-ligand complex;
(f) providing a second probe, comprising a second fluorescent
metal-ligand complex coupled to a second oligonucleotide
having a sequence complimentary to a second fragment of said
strand differing from said first fragment by at least one base;
(g) adding said second probe to a sample that contains said second
fragment to form a second mixture containing a second reaction
product of said second probe and said second fragment;
(h) exposing said second mixture to an exciting amount of
radiation;
(i) detecting fluorescence of said second metal-ligand complex;
(j) identifying a second base sequence of said second fragment
based on fluorescence of said second metal-ligand complex;
(k) comparing said second base sequence with said first base
sequence to identify a difference between the first and second
sequences and thereby determine a base sequence of said
nucleotide strand;
complex is selected from the group consisting of Co, Cr, Cu,
Mo, Rh, W, Re, Os, Ir, and Pt;
wherein said detection utilizes measurement of fluorescence
lifetime; and
wherein autofluorescence is suppressed by fluorescence gating.
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