Patent ID: 6150107
Filing Date: 2000-11-21
Classification: C12Q

Abstract:
In a method for sequencing a nucleic acid fragment, comprising:(i) hybridizing an oligonucleotide primer to said fragment,(ii) copying said fragment with a DNA polymerase in separate reaction vessels, each reaction vessel containing dNTPs, one of a set of distinct labeled oligonucleotide primers, all of which bind to a complementary sequence in said fragment, and one of a plurality of dideoxynucleotides, to generate single stranded DNA sequencing fragments, each labeled oligonucleotide primer comprising an oligonucleotide backbone having bonded thereto a donor fluorophore and an acceptor fluorophore in energy transfer relationship, the donor fluorophore of each labeled oligonucleotide primer absorbing light at a wavelength that is substantially common among all labeled oligonucleotide primers and the acceptor fluorophores emitting light energy at wavelengths that differ among labeled oligonucleotide primers in said set, and(iii) separating the resulting mixture of single-stranded DNA sequencing fragments into bands and determining the sequence by means of the order of the bands, the improvement in which the donor fluorophore of each labeled oligonucleotide primer is a cyanine dye characterized by a quantum yield of between about 0.01 and about 0.25, and each acceptor fluorophore has a quantum yield of at least 0.3.