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Instrument and method for the sequencing of genome

  • US 5,288,644 A
  • Filed: 11/13/1992
  • Issued: 02/22/1994
  • Est. Priority Date: 04/04/1990
  • Status: Expired due to Term
First Claim
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1. A method of DNA sequencing of the genome by detecting the mass weights of DNA fragments comprising the steps of:

  • (a) producing a piece of DNA of unknown sequence;

    (b) performing four different base-specific reactions on the piece of DNA to produce four different DNA sets containing DNA fragments each DNA set having a common origin and terminating at a particular base along the unknown sequence;

    (c) selecting a solid matrix having a strong absorption band at the wavelength of light produced by a laser and placing one of the DNA sets on the matrix, the strong absorption band of the matrix having a longer wavelength than the absorption bands for the DNA set;

    (d) without separation of the DNA fragments by electrophoresis or other separation methods and without labeling the DNA fragments with radioactive, fluorescent or other labeling means, striking the DNA fragments on the solid matrix within one of the DNA sets with a series of laser pulses from the laser to desorb ions of the DNA fragments and produce ionized DNA fragments;

    (e) detecting the mass weights of the ionized DNA fragments by a time of flight mass spectrometer; and

    (f) repeating steps (d) and (e) for the other DNA sets to determine the sequence of bases in the DNA.

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