Patent ID: 9677067
Date: 2017-06-13
CPC Classifications: C12N,C12P

Claim:
1. A method for nucleic acid assembly, the method comprising: a) providing a predetermined nucleic acid sequence; b) providing a plurality of precursor double-stranded nucleic acid fragments, each precursor double-stranded nucleic acid fragment having two strands, wherein each of the two strands comprises a sticky end sequence of 5′-A (N c) providing primers comprising a nicking endonuclease recognition site and a sequence comprising (i) 5′-A (N d) performing a polynucleotide extension reaction to form a polynucleotide extension reaction product that is double-stranded nucleic acid fragments; e) subjecting the polynucleotide extension reaction product to nicking and cleavage reactions to form double-stranded nucleic acid fragments with 3′ overhangs; and f) annealing the double-stranded nucleic acid fragments with 3′ overhangs to form a nucleic acid encoding for the predetermined nucleic acid sequence that does not include the nicking endonuclease recognition site.