A process for the electrophoretic analysis of DNA fragments produced in DNA sequencing operations wherein chromophores or fluorophores are used to tag the DNA fragments produced by the sequencing chemistry and permit the detection and characterization of the fragments as they are resolved by electrophoresis through a gel. Preferably four different fragment sets are tagged with the fluorophores fluorescein, Texas Red, tetramethyl rhodamine, and 7-nitro-benzofurazan. A system for the electrophoretic analysis of DNA fragments produced in DNA sequencing operations comprising: a source of chromophore or fluorescent tagged DNA fragments; a zone for contacting an electrophoresis gel; means for introducing said tagged DNA fragments to said zone; and photometric means for monitoring said tagged DNA fragments as they move through said gel.

Claims

56 · 6 independent · depth 4

56 granted claims

›1.independentA method of separating and detecting tagged polynucleotides which comprises: providing a plurality of polynucleotides, each tagged with a chromophore …+5

›7.independentA method of detecting a tagged polynucleotide, which comprises: providing a plurality of polynucleotides, wherein each of said plurality of polynucleo…+6

›8.↳ 7The method according to claim 7, wherein the tagged polynucleotides are obtained from one of a set of primer extension reactions in which each of the …d2

›9.↳ 7The method according to claim 7, wherein prior to electrophoresis, each primer oligonucleotide is coupled to the chromophore or fluorophore by an amin…d2

›10.↳ 7The method according to claim 7, wherein the primer oligonucleotide is tagged with the chromophore or fluorophore to allow chain extension by a polyme…d2

›11.↳ 7The method according to claim 7, wherein the primer oligonucleotide is tagged such that the primer oligonucleotide has a free 3' hydroxyl group.d2

›12.↳ 7The method of claim 7, wherein the detecting of the resolved tagged polynucleotides comprises exposure of the tagged polynucleotides to a high-intensi…d2

›13.↳ 7The method of claim 7, wherein the detecting of the resolved tagged polynucleotides comprises exposure of the tagged polynucleotides to a laser.d2

›14.independentA method of determining the sequence of a polynucleotide by analyzing polynucleotide fragments generated by a polynucleotide sequencing technique, eac…+13

›15.↳ 14The method according to claim 14, wherein the polynucleotide is DNA.d2

›16.↳ 14The method according to claim 14, wherein each tagged polynucleotide fragment comprises an identical primer oligonucleotide sequence, and the primer o…d2+2

›19.↳ 14The method according to claim 14, wherein the method is a chemical degradation method and the polynucleotide fragments are tagged with the chromophore…d2

›20.↳ 14The method according to claim 14, wherein the tagged fragments are obtained from a primer extension reaction in which the primer is tagged with a chro…d2

›21.↳ 14The method according to claim 14, wherein the tagged fragments from at least two of the sequencing reactions A, C, G and T are distinguishable from on…d2

›22.↳ 14The method according to claim 14, wherein the tagged fragments from all of the sequencing reactions A, C, G and T are distinguishable from one another…d2

›23.↳ 14The method according to claim 14, wherein prior to electrophoresis, at least one of the polynucleotide fragments is coupled to the chromophore or fluo…d2

›24.↳ 14The method according to claim 14, wherein the step of detecting the polynucleotide fragments is performed during the electrophoresis.d2

›25.↳ 14The method according to claim 14, wherein the polynucleotide sequencing technique comprises a Maxam/Gilbert chemical degradation reaction or a Sanger …d2

›26.↳ 14The method of claim 14, wherein the detecting of the separated tagged polynucleotide fragments comprises exposure of the tagged polynucleotide to a hi…d2

›27.↳ 14The method of claim 14, wherein the detecting of the separated tagged polynucleotide comprises exposure of the tagged polynucleotide to a laser.d2

›28.independentA method for determining the sequence of a polynucleotide which comprises: providing polynucleotide fragments tagged with chromophores or fluorophores…+12

›29.↳ 28The method according to claim 28 wherein the polynucleotide is DNA.d2

›30.↳ 28The method according to claim 28, wherein each tagged polynucleotide fragment comprises an identical primer oligonucleotide sequence, and the primer o…d2

›31.↳ 28The method according to claim 28, wherein the method is a chemical degradation method and the polynucleotide fragments are labeled with the chromophor…d2

›32.↳ 28The method according to claim 28, wherein the tagged fragments are obtained from a primer extension reaction in which the primer is tagged with a chro…d2

›33.↳ 28The method according to claim 28, wherein the tagged fragments from at least two of the sequencing reactions A, C, G and T are distinguishable from on…d2

›34.↳ 28The method according to claim 28, wherein the tagged fragments from all of the sequencing reactions A, C, G and T are distinguishable from one another…d2+1

›36.↳ 28The method according to claim 28, wherein prior to electrophoresis, at least one of the polynucleotide fragments are coupled to the chromophore or flu…d2

›37.↳ 28The method according to claim 28, wherein one or more polynucleotide fragments are provided with a protected amino group prior to electrophoresis, whi…d2+1

›39.↳ 28The method of claim 28, wherein the detecting of the resolved fragments comprises exposure of the resolved fragments to a high-intensity monochromatic…d2

›40.↳ 28The method of claim 28, wherein the detecting of the resolved fragments comprises exposure of the resolved fragments to a laser.d2

›41.independentA method for determining the sequence of a polynucleotide which comprises: providing polynucleotide fragments generated by a polynucleotide sequencing…+12

›42.↳ 41The method according to claim 41 wherein the polynucleotide is DNA.d2

›43.↳ 41The method according to claim 41, wherein each tagged polynucleotide fragment comprises an identical primer oligonucleotide sequence, and the primer o…d2

›44.↳ 41The method according to claim 41, wherein the method is a chemical degradation method and the polynucleotide fragments are labeled with the chromophor…d2

›45.↳ 41The method according to claim 41, wherein the tagged fragments are obtained from a primer extension reaction in which the primer is tagged with a chro…d2

›46.↳ 41The method according to claim 41, wherein the tagged fragments from at least two of the sequencing reactions A, C, G and T are distinguishable from on…d2

›47.↳ 41The method according to claim 41, wherein the tagged fragments from all of the sequencing reactions A, C, G and T are distinguishable from one another…d2+1

›49.↳ 41The method according to claim 41, wherein the fragments from one or more of four sequencing reactions A, C, G or T are coupled to the chromophore or f…d2+1

›51.↳ 41The method according to claim 41, wherein the fragments from one or more of four sequencing reactions A, C, G or T are provided with a protected amino…d2

›52.↳ 41The method of claim 41, wherein the detecting of the fragments as they are being resolved comprises exposure of the fragments to a high-intensity mono…d2

›53.↳ 41The method of claim 41, wherein the detecting of the fragments as they are being resolved comprises exposure of the fragments to a laser.d2

›54.independentA method of determining the sequence of a single-stranded polynucleotide comprising: obtaining a set of fragments of the complement of said single-str…+2

Classifications

6 codes

USPC · US Patent Classification

435/6530/25.31435/91.1530/25.32530/24.3530/25.3

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Stephanie W. Zitomer

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