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DNA sequencing by mass spectrometry via exonuclease degradation

  • US 6,074,823 A
  • Filed: 11/06/1996
  • Issued: 06/13/2000
  • Est. Priority Date: 03/19/1993
  • Status: Expired due to Term
First Claim
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1. A process for determining the sequence of a target nucleic acid, comprising the steps of:

  • (i) obtaining multiple nucleic acid copies of the target nucleic acid, wherein the multiple copies contain at least one mass-modified nucleotide, corresponding to one of the four possible nucleotide bases;

    (ii) cleaving the multiple nucleic acid copies from a first end to a second end with an exonuclease having an activity, which is inhibited by the mass-modified nucleotide, thereby generating base-terminated nucleic acid fragments;

    (iii) identifying the nested nucleic acid fragments by mass spectrometry; and

    (iv) determining the sequence of the target nucleic acid from the identified nested nucleic acid fragment.

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