Skip to main content
SpringerLink
Log in

BEAMing: single-molecule PCR on microparticles in water-in-oil emulsions

  • Protocol
  • Published:

From Nature Methods

View current issue Submit your manuscript

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2: Phase contrast micrograph (viewed at 400×) of emulsions deposited in the well of a 48-well tissue culture plate.
Figure 3: Analysis of bead populations with flow cytometry.

References

  1. Cohen, S.N., Chang, A.C., Boyer, H.W. & Helling, R.B. Construction of biologically functional bacterial plasmids in vitro. Proc. Natl. Acad. Sci. USA 70, 3240–3244 (1973).

    Article  CAS  Google Scholar 

  2. Saiki, R.K. et al. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 230, 1350–1354 (1985).

    Article  CAS  Google Scholar 

  3. Tawfik, D.S. & Griffiths, A.D. Man-made cell-like compartments for molecular evolution. Nat. Biotechnol. 16, 652–656 (1998).

    Article  CAS  Google Scholar 

  4. Dressman, D., Yan, H., Traverso, G., Kinzler, K.W. & Vogelstein, B. Transforming single DNA molecules into fluorescent magnetic particles for detection and enumeration of genetic variations. Proc. Natl. Acad. Sci. USA 100, 8817–8822 (2003).

    Article  CAS  Google Scholar 

  5. Diehl, F. et al. Detection and quantification of mutations in the plasma of patients with colorectal tumors. Proc. Natl. Acad. Sci. USA 102, 16368–16373 (2005).

    Article  CAS  Google Scholar 

  6. Li, M., Diehl, F., Dressman, D., Vogelstein, B. & Kinzler, K.W. BEAMing up for detection and quantification of rare sequence variants. Nat. Methods 3, 95–97 (2006).

    Article  CAS  Google Scholar 

  7. Shendure, J. et al. Accurate multiplex polony sequencing of an evolved bacterial genome. Science 309, 1728–1732 (2005).

    Article  CAS  Google Scholar 

  8. Mercier, J.F. & Slater, G.W. Solid phase DNA amplification: a Brownian dynamics study of crowding effects. Biophys. J. 89, 32–42 (2005).

    Article  CAS  Google Scholar 

  9. Utada, A.S. et al. Monodisperse double emulsions generated from a microcapillary device. Science 308, 537–541 (2005).

    Article  CAS  Google Scholar 

  10. Nagai, H., Murakami, Y., Morita, Y., Yokoyama, K. & Tamiya, E. Development of a microchamber array for picoliter PCR. Anal. Chem. 73, 1043–1047 (2001).

    Article  CAS  Google Scholar 

  11. Aharoni, A. et al. High-throughput screening of enzyme libraries: new thiobutyrolactonases by fluorescence activated sorting of single cells in emulsion compartments. Chem. Biol. 12, 1281–1289 (2005).

    Article  CAS  Google Scholar 

  12. Mastrobattista, E. et al. High-throughput screening of enzyme libraries: in vitro evolution of a β-galactosidase by fluorescence-activated sorting of double emulsions. Chem. Biol. 12, 1291–1300 (2005).

    Article  CAS  Google Scholar 

  13. Margulies, M. et al. Genome sequencing in microfabricated high-density picolitre reactors. Nature 437, 376–380 (2005).

    Article  CAS  Google Scholar 

  14. Kojima, T. et al. PCR amplification from single DNA molecules on magnetic beads in emulsion: application for high-throughput screening of transcription factor targets. Nucleic Acids Res. 33, e150 (2005).

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. The Howard Hughes Medical Institute, and The Ludwig Center for Cancer Genetics and Therapeutics, The Johns Hopkins Kimmel Cancer Center, 1650 Orleans Street, Baltimore, 21231, Maryland, USA

    Frank Diehl, Meng Li, Yiping He, Kenneth W Kinzler, Bert Vogelstein & Devin Dressman

Authors
  1. Frank Diehl
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Meng Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yiping He
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kenneth W Kinzler
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Bert Vogelstein
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Devin Dressman
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Bert Vogelstein.

Ethics declarations

Competing interests

Under a licensing agreement between EXACT Sciences and The Jonhs Hopkins University, K.W.K. and B.V. are entitled to a share of royalties received by the university on sales of products related to digital PCR. Under a licensing agreement between Agencourt Biosciences Corporation and The Jonhs Hopkins University, D.D., K.W.K. and B.V. are entitled to a share of royalties received by the university on sales of products related to the use of BEAMing for preparing templates for DNA sequencing. The terms of these arrangements are being managed by The Jonhs Hopkins University in accordance with its conflict of interest policies.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Diehl, F., Li, M., He, Y. et al. BEAMing: single-molecule PCR on microparticles in water-in-oil emulsions. Nat Methods 3, 551–559 (2006). https://doi.org/10.1038/nmeth898

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmeth898

  • Springer Nature America, Inc.

This article is cited by

Search

Navigation