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End-complementary polymerase reaction

  • US 5,928,905 A
  • Filed: 07/03/1996
  • Issued: 07/27/1999
  • Est. Priority Date: 04/18/1995
  • Status: Expired due to Term
First Claim
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1. A method for amplifying a target polynucleotide, comprising(1) providing a series of overlapping polynucleotides of alternating polarity spanning the target polynucleotide, wherein each given polynucleotide in the series has two flanking polynucleotides, and the 3'"'"' end of the given polynucleotide is complementary to the 3'"'"' end of one flanking polynucleotide and the 5'"'"' end of the given polynucleotide is complementary to the 5'"'"' end of the other flanking polynucleotide;

  • (2) incubating the polynucleotides under conditions whereby complementary ends anneal;

    (3) extending the 3'"'"' ends of annealed polynucleotides with a polymerase, under conditions whereby each polynucleotide serves as a template for another;

    (4) denaturing the extended polynucleotides; and

    (5) repeating steps (2)-(4) 1-100 times, whereby the extended polynucleotides in step (4) constitute the polynucleotides in step (2) in the next cycle.

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