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Massive parallel method for decoding DNA and RNA

DC CAFC
  • US 8,088,575 B2
  • Filed: 07/19/2010
  • Issued: 01/03/2012
  • Est. Priority Date: 10/06/2000
  • Status: Expired due to Fees
First Claim
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1. A method of determining the identity of a nucleotide analogue incorporated into a nucleic acid primer extension strand, comprising:

  • a) contacting a nucleic acid template attached to a solid surface with a nucleic acid primer which hybridizes to the template;

    b) simultaneously contacting the product of step a) with a polymerase and four nucleotide analogues which are either (i) aA, aC, aG, and aT, or (ii) aA, aC, aG, and aU, so as to incorporate one of the nucleotide analogues onto the nucleic acid primer and form a nucleic acid primer extension strand, wherein each nucleotide analogue within (i) or (ii) comprises a base labeled with a unique label and contains a small removable chemical moiety capping the 3′

    -OH group of the sugar of the nucleotide analogue, wherein said small cleavable chemical group does not interfere with the recognition of the nucleotide analogue by polymerase as a substrate; and

    c) detecting the unique label of the incorporated nucleotide analogue, so as to thereby determine the identity of the nucleotide analogue incorporated into the nucleic acid primer extension strand.

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