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DNA sequencing by mass spectrometry via exonuclease degradation

  • US 5,851,765 A
  • Filed: 05/30/1995
  • Issued: 12/22/1998
  • Est. Priority Date: 03/19/1993
  • Status: Expired due to Term
First Claim
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1. A multiplex mass spectrometric method for simultaneously determining the sequence of at least two different nucleic acids concurrently, comprising the steps of:

  • (i) obtaining multiple copies of the at least two different nucleic acids, wherein at least one of the at least two different nucleic acids comprises mass modified nucleotides;

    (ii) concurrently cleaving the at least two different nucleic acids unilaterally from a first end to a second end with an exonuclease to generate sequentially released individual nucleotides and optionally, at least two different sets of nucleic acid fragments;

    (iii) identifying each of the sequentially released nucleotides by mass spectrometry or determining the molecular weight values of each of the sets of nucleic acid fragments by mass spectrometry; and

    (iv) determining the sequence of the at least two different nucleic acids based on the molecular weight values of the sequentially released individual nucleotides or the sets of nucleic acid fragments.

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