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Nucleic acid target generation

  • US 5,270,184 A
  • Filed: 11/19/1991
  • Issued: 12/14/1993
  • Est. Priority Date: 11/19/1991
  • Status: Expired due to Term
First Claim
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1. A method for generating an amplifying a nucleic acid fragment which comprises:

  • (a) specifically hybridizing a first primer 5'"'"' to target nucleic acid sequence, the first primer containing a restriction enzyme recognition sequence 5'"'"' to a target binding region,(b) simultaneously with (a), hybridizing a second primer 5'"'"' to the first primer,(c) extending the first and second primer so that the first primer extension product is displaced from the target nucleic acid sequence by extension of the second primer,(d) making the first primer extension product double stranded by synthesizing a complementary strand, and(e) amplifying the first primer extension product in an amplification reaction in which the restriction enzyme recognition site of the double stranded nucleic acid fragment is nicked by a restriction enzyme.

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