Enzyme Engineering

Volume 978 of the series Methods in Molecular Biology pp 165-171


Gene Synthesis by Assembly of Deoxyuridine-Containing Oligonucleotides

  • Romualdas VaisvilaAffiliated withNew England Biolabs, Inc. Email author 
  • , Jurate BitinaiteAffiliated withNew England Biolabs, Inc.

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Gene synthesis is an invaluable technique in synthetic and molecular biology for synthesis of artificial genes, operons, and even genomes. In many cases the traditional methods for obtaining functional DNA sequences through cloning are not applicable due to the novelty of genetic material. Here, we describe the simple and economical DNA synthesis method based on USER™ technology. The method consists of (1) synthesis of building blocks up to 500 bp; (2) assembly of genes up to 3 kb; (3) error correction reassembly; and (4) assembly of operons up to 15 kb if needed.

Key words

Gene synthesis Polymerase chain reaction Uracil excision USER cloning Synthetic biology