Skip to main content

Using DNAWorks in Designing Oligonucleotides for PCR-Based Gene Synthesis

  • Protocol
  • First Online:
Gene Synthesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 852))

Abstract

The availability of sequences of entire genomes has dramatically increased the number of protein targets, many of which will need to be overexpressed in cells other than where they have been identified originally. Gene synthesis often provides a fast and economically efficient approach. The synthetic gene can be optimized for expression and constructed for easy mutational manipulation without regard for the parent genome. Yet design and construction of synthetic genes, especially those coding for large proteins, can be a slow, difficult, and confusing process. DNAWorks automates the design of oligonucleotides for gene synthesis by PCR-based methods.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Carr PA, Church GM (2009) Genome engineering. Nature Biotech 27:1151–1162.

    Article  CAS  Google Scholar 

  2. Prodromou C, Pearl LH (1992) Recursive PCR: a novel technique for total gene synthesis. Protein Eng 5:827–829.

    Article  PubMed  CAS  Google Scholar 

  3. Dillon PJ, Rosen CA (1990) A rapid method for the construction of synthetic genes using the polymerase chain reaction. Biotechniques 9:298–300.

    PubMed  CAS  Google Scholar 

  4. Gao X, Yo P et al. (2003) Thermodynamically balanced inside-out (TBIO) PCR-based gene synthesis: a novel method of primer design for high-fidelity assembly of longer gene sequences. Nucleic Acids Res 31:e143.

    Article  PubMed  Google Scholar 

  5. Czar MJ, Anderson JC et al. (2009) Gene synthesis demystified. Trends Biotechnol 27:63–72.

    Article  PubMed  CAS  Google Scholar 

  6. Xiong AS, Peng R et al. (2008) Chemical gene synthesis: strategies, softwares, error corrections, and applications. FEMS Microbiol Rev 32:522–540.

    Article  PubMed  CAS  Google Scholar 

  7. Hoover DM, Lubkowski J (2002) DNAWorks: an automated method for designing oligonucleotides for PCR-based gene synthesis. Nucleic Acids Res 30:e43.

    Article  PubMed  Google Scholar 

  8. Nakamura Y et al. (2000) Nucleic Acids Res 28:292.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to David Hoover .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Sceince+Business Media, LLC

About this protocol

Cite this protocol

Hoover, D. (2012). Using DNAWorks in Designing Oligonucleotides for PCR-Based Gene Synthesis. In: Peccoud, J. (eds) Gene Synthesis. Methods in Molecular Biology, vol 852. Humana Press. https://doi.org/10.1007/978-1-61779-564-0_16

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-564-0_16

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-563-3

  • Online ISBN: 978-1-61779-564-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics