Unit

Gene Assembly from Chip-Synthesized Oligonucleotides

  1. Nikolai Eroshenko1,5,
  2. Sriram Kosuri2,3,5,
  3. Adam H. Marblestone3,4,
  4. Nicholas Conway3,
  5. George M. Church2,3

Published Online: 1 MAR 2012

DOI: 10.1002/9780470559277.ch110190

Current Protocols in Chemical Biology

Current Protocols in Chemical Biology

How to Cite

Eroshenko, N., Kosuri, S., Marblestone, A. H., Conway, N. and Church, G. M. 2012. Gene Assembly from Chip-Synthesized Oligonucleotides. Current Protocols in Chemical Biology. 4:1–17.

Author Information

  1. 1

    Harvard School of Engineering and Applied Sciences, Cambridge, Massachusetts

  2. 2

    Department of Genetics, Harvard Medical School, Boston, Massachusetts

  3. 3

    Wyss Institute for Biologically Inspired Engineering, Boston, Massachusetts

  4. 4

    Harvard Biophysics Program, Cambridge, Massachusetts

  5. 5

    These authors contributed equally to this work

Publication History

  1. Published Online: 1 MAR 2012
  2. Published Print: MAR 2012

Abstract

De novo synthesis of long double-stranded DNA constructs has a myriad of applications in biology and biological engineering. However, its widespread adoption has been hindered by high costs. Cost can be significantly reduced by using oligonucleotides synthesized on high-density DNA chips. However, most methods for using off-chip DNA for gene synthesis have failed to scale due to the high error rates, low yields, and high chemical complexity of the chip-synthesized oligonucleotides. We have recently demonstrated that some commercial DNA chip manufacturers have improved error rates, and that the issues of chemical complexity and low yields can be solved by using barcoded primers to accurately and efficiently amplify subpools of oligonucleotides. This unit includes protocols for computationally designing the DNA chip, amplifying the oligonucleotide subpools, and assembling 500- to 800-bp constructs. Curr. Protoc. Chem. Biol. 4:1-17 © 2012 by John Wiley & Sons, Inc.

Keywords:

  • oligonucleotide;
  • gene synthesis;
  • nucleic acids;
  • synthetic biology