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Multiple tag labeling method for DNA sequencing

  • US 5,436,130 A
  • Filed: 10/26/1993
  • Issued: 07/25/1995
  • Est. Priority Date: 03/19/1992
  • Status: Expired due to Fees
First Claim
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1. A method of determining the sequence of DNA which comprises the steps ofprocessing said DNA to form four sets of DNA sequencing fragments where one set contains fragments terminating in G, a second set contains fragments terminating in A, a third set contains fragments terminating in T, and a fourth set contains fragments terminating in C, such that selected mole fractions of each one of selected sets of sequencing fragments is tagged with a first fluorophore which fluoresces at a first wavelength and such that different selected mole fractions of each one of selected sets of sequencing fragments is tagged with a second fluorophore which fluoresces at a second wavelength and in which one of said sets of sequencing fragments is comprised of a mixture of selected mole fractions of fragments where one mole fraction is labeled with the first fluorophore and the other mole fraction is labeled with the second fluorophore,separating said sets of tagged DNA sequencing fragments in a single channel or lane, anddetermining the sequence of said sequencing fragments by detecting the ratio of fluorescence intensity at the first and second wavelengths as a function of time or position.

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