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Sequencing by incorporation

  • US 6,613,513 B1
  • Filed: 02/22/2000
  • Issued: 09/02/2003
  • Est. Priority Date: 02/23/1999
  • Status: Expired due to Term
First Claim
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1. A method of sequencing a nucleic acid, the method comprising:

  • (i) providing a nucleic acid template and a primer;

    (ii) incubating the nucleic acid template and primer with a polymerase and one or more fluorescently labeled nucleotides, thereby adding at least one of the one or more fluorescently labeled nucleotides to the primer, resulting in an extended primer;

    (iii) detecting the at least one fluorescently labeled nucleotide added to the primer;

    (iv) photobleaching the at least one or the one or more fluoroescently labeled nucleotides on the extended primer by applying a light pulse, wherein photobleaching destroys or reduces the fluorescence of the at least one of the one or more fluorescently labeled nucleotides to an acceptable level without removing the labeled nucleotide(s) from the extended primer.

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