Gene Synthesis by Assembly of Deoxyuridine-Containing Oligonucleotides

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 978)

Abstract

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 

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Copyright information

© Springer New York 2013

Authors and Affiliations

  1. 1.New England Biolabs, Inc.IpswichUSA

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