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Methods for recombinatorial nucleic acid synthesis

  • US 6,994,963 B1
  • Filed: 07/10/2000
  • Issued: 02/07/2006
  • Est. Priority Date: 07/10/2000
  • Status: Active Grant
First Claim
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1. A method for creating a plurality of nucleic acids comprising the steps of:

  • (a) annealing a plurality of a defined first primer nucleic acid to a plurality of at least one first single stranded template nucleic acid,(b) performing a first extension by extending the first primer nucleic acid employing the first template nucleic acid in the presence of at least one dideoxynucleotide or a dideoxynucleotide comprising a nucleotide analog, wherein the dideoxynucleotide or dideoxynucleotide comprising a nucleotide analog is incorporated into the extended nucleic acid, to form a plurality of first extended nucleic acids having essentially identical 5′

    ends and variable 3′

    ends,(c) denaturing the first extended nucleic acids from the first template nucleic acid,(d) modifying or removing the dideoxynucleotide or dideoxynucleotide comprising a nucleotide analog from the first extended nucleic acids,e) annealing the first extended nucleic acids to a plurality of at least a second single stranded template nucleic acid whose sequence is not identical to the first template nucleic acid, andf) performing a second extension by extending the extended nucleic acid employing the second template nucleic acid to form a plurality of twice extended nucleic acids having essentially identical 5′

    ends.

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