Skip to main content

Amplification of Fungal Genomes Using Multiple Displacement Amplification

  • Protocol
  • First Online:
Molecular and Cell Biology Methods for Fungi

Part of the book series: Methods in Molecular Biology ((MIMB,volume 638))

  • 4494 Accesses

Abstract

The availability of genomic DNA of sufficient quality and quantity is fundamental to molecular genetic analysis. Many filamentous fungi are slow growing or even unculturable and current DNA isolation methods are often unsatisfactory. Multiple displacement amplification (MDA) is a technique that can be employed to reliably amplify whole genomes from such recalcitrant species. Template DNA obtained using traditional DNA extraction methods, glass bead-mediated disruption of fungal spores or alkaline lysis of mycelium can be used to produce DNA of sufficient quality to be used as a substrate in MDA. With the advent of next generation sequencing methods, the ability to utilize relatively small samples of DNA to achieve complete genome sequencing is now a possibility.

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. Foster SJ, Monahan BJ, Bradshaw RE (2006) Genomics of the filamentous fungi - moving from the shadow of the bakers yeast. Mycologist 20:10–14

    Article  Google Scholar 

  2. Al-Samarrai TH, Schmid J (2000) A simple method for extraction of fungal genomic DNA. Lett Appl Microbiol 30:53–56

    Article  CAS  PubMed  Google Scholar 

  3. Foster SJ, Monahan BJ (2005) Whole genome amplification from filamentous fungi using Phi29-mediated multiple displacement amplification (MDA). Fungal Genet Biol 42:367–375

    Article  CAS  PubMed  Google Scholar 

  4. Dean FB, Hosono S, Fang LH, Wu XH, Faruqi AF, Bray-Ward P, Sun ZY, Zong QL, Du YF, Du J, Driscoll M, Song WM, Kingsmore SF, Egholm M, Lasken RS (2002) Comprehensive human genome amplification using multiple displacement amplification. Proc Natl Acad Sci U S A 99:5261–5266

    Article  CAS  PubMed  Google Scholar 

  5. Wang Y, Zhu M, Zhang R, Yang H, Wang Y, Sun G, Jin S, Hsiang T (2009) Whole genome amplification of the rust Puccinia striiformis f. sp. tritici from single spores. J Microbiol Methods 77:229-234. doi:10.1016/j.mimet.2009.02.007

    Article  CAS  PubMed  Google Scholar 

  6. Telenius H, Carter NP, Bebb CE, Nordenskjold M, Ponder BAJ, Tunnacliffe A (1992) Degenerate oligonucleotide-primed PCR - general amplification of target DNA by a single degenerate primer. Genomics 13:718–725

    Article  CAS  PubMed  Google Scholar 

  7. Zhang L, Cui XF, Schmitt K, Hubert R, Navidi W, Arnheim N (1992) Whole genome amplification from a single cell - implications for genetic analysis. Proc Natl Acad Sci U S A 89:5847–5851

    Article  CAS  PubMed  Google Scholar 

  8. Lasken RS, Egholm M (2003) Whole genome amplification: abundant supplies of DNA from precious samples or clinical specimens. Trends Biotechnol 21:531–535

    Article  CAS  PubMed  Google Scholar 

  9. Esteban JA, Salas M, Blanco L (1993) Fidelity of Phi 29 DNA polymerase Comparison between protein-primed initiation and DNA polymerization. J Biol Chem 268:2719–2726

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We wish to thank Prof. Barry Scott, Massey University, New Zealand who granted us the time and materials to conduct this work in his laboratory.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Simon J. Foster .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2010 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Foster, S.J., Monahan, B.J. (2010). Amplification of Fungal Genomes Using Multiple Displacement Amplification. In: Sharon, A. (eds) Molecular and Cell Biology Methods for Fungi. Methods in Molecular Biology, vol 638. Humana Press. https://doi.org/10.1007/978-1-60761-611-5_13

Download citation

  • DOI: https://doi.org/10.1007/978-1-60761-611-5_13

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-610-8

  • Online ISBN: 978-1-60761-611-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics