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Method, apparatus and kits for sequencing of nucleic acids using multiple dyes

  • US 20030003498A1
  • Filed: 08/12/2002
  • Published: 01/02/2003
  • Est. Priority Date: 04/18/1996
  • Status: Abandoned Application
First Claim
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1. A method for evaluating the nucleic acid sequence of a plurality of samples comprising the steps of (a) obtaining a first aliquot of each sample;

  • (b) combining the first aliquot of each sample with a sequencing reaction mixture containing a polymerase enzyme, a primer for hybridizing with the sample, nucleoside feedstocks and a first dideoxynucleoside to form a first plurality of mixtures of product oligonucleotide fragments, one for each sample, wherein the product oligonucleotide fragments formed from each first aliquot are labeled with different fluorescent tags, said different fluorescent tags being distinguishable one from the other on the basis of their excitation or emission spectra;

    (c) combining the first plurality of mixtures of oligonucleotide products to form a first combined mixture;

    (d) loading the first combined mixture onto a separation matrix at a first loading site;

    (e) applying an electric field to cause the oligonucleotide products to migrate within the separation matrix; and

    (f) detecting the oligonucleotide products having the different fluorescent tags as they migrate within the separation matrix.

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