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Isothermal DNA amplification

  • US 20060166250A1
  • Filed: 01/23/2006
  • Published: 07/27/2006
  • Est. Priority Date: 01/25/2005
  • Status: Active Grant
First Claim
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1. A method of amplifying a polynucleotide comprising the steps of:

  • providing a single stranded DNA containing a promoter sequence and a polynucleotide, the single stranded DNA having a 5′

    end, a 3′

    end, and complementary sequences at the 3′

    end capable of forming a hairpin having a loop and a duplex, the promoter sequence being disposed between the polynucleotide and the 3′

    end and the single stranded DNA being attached to a solid phase support by its 5′

    end;

    extending the duplex to form a double stranded DNA that includes the polynucleotide and complement thereof and the promoter sequence and complement thereof, thereby forming an operational promoter; and

    generating RNA copies of the polynucleotide with an RNA polymerase that recognizes the operational promoter.

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