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High-fidelity DNA sequencing using solid phase capturable dideoxynucleotides and mass spectrometry

  • US 20030027140A1
  • Filed: 03/30/2001
  • Published: 02/06/2003
  • Est. Priority Date: 03/30/2001
  • Status: Abandoned Application
First Claim
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1. A method for sequencing DNA by detecting the identity of a dideoxynucleotide incorporated to the 3′

  • end of a DNA sequencing fragment using mass spectrometry, which comprises;

    (a) attaching a chemical moiety via a linker to a dideoxynucleotide to produce a labeled dideoxynucleotide;

    (b) terminating a DNA sequencing reaction with the labeled dideoxynucleotide to generate a labeled DNA sequencing fragment, wherein the DNA sequencing fragment has a 3′

    end and the chemical moiety is attached via the linker to the 3′

    end of the DNA sequencing fragment;

    (c) capturing the labeled DNA sequencing fragment on a surface coated with a compound that specifically interacts with the chemical moiety attached via the linker to the DNA sequencing fragment, thereby capturing the DNA sequencing fragment;

    (d) washing the surface to remove any non-bound component;

    (e) freeing the DNA sequencing fragment from the surface; and

    (f) analyzing the DNA sequencing fragment using mass spectrometry so as to sequence the DNA.

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