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MegaPlex PCR: a strategy for multiplex amplification

Abstract

'MegaPlex PCR' is a robust technology for highly multiplexed amplification of specific DNA sequences. It uses target-specific pairs of PCR primers that are physically separated by surface immobilization. Initial surface-based amplification cycles are then coupled to efficient solution-phase PCR using one common primer pair. We demonstrate this method by co-amplifying and genotyping 75 unselected human single-nucleotide polymorphism (SNP) loci.

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Figure 1: Schematic representation of MegaPlex PCR.
Figure 2: Analysis of the MP50 and MP75 amplification products.

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References

  1. Markoulatos, P., Siafakas, N. & Moncany, M. J. Clin. Lab. Anal. 16, 47–51 (2002).

    Article  CAS  Google Scholar 

  2. Vallone, P.M. & Butler, J.M. Biotechniques 37, 226–231 (2004).

    Article  CAS  Google Scholar 

  3. Wang, H.Y. et al. Genome Res. 15, 276–283 (2005).

    Article  CAS  Google Scholar 

  4. Adessi, C. et al. Nucleic Acids Res. 28, e87 (2000).

    Article  CAS  Google Scholar 

  5. Shapero, M.H., Leuther, K.K., Nguyen, A., Scott, M. & Jones, K.W. Genome Res. 11, 1926–1934 (2001).

    Article  CAS  Google Scholar 

  6. Pemov, A., Modi, H., Chandler, D.P. & Bavykin, S. Nucleic Acids Res. 33, e11 (2005).

    Article  CAS  Google Scholar 

  7. Brownie, J. et al. Nucleic Acids Res. 25, 3235–3241 (1997).

    Article  CAS  Google Scholar 

  8. Shapero, M.H. et al. Nucleic Acids Res. 32, e181 (2004).

    Article  Google Scholar 

  9. Eichinger, L. et al. Nature 435, 43–57 (2005).

    Article  CAS  Google Scholar 

  10. Fredriksson, S. et al. Nucleic Acids Res. 35, e47 (2007).

    Article  Google Scholar 

  11. Hansen, A. et al. Nat. Methods 4, 35–37 (2007).

    Article  CAS  Google Scholar 

  12. Wang, D.G. et al. Science 280, 1077–1082 (1998).

    Article  CAS  Google Scholar 

  13. Baum, M. et al. Nucleic Acids Res. 31, e151 (2003).

    Article  Google Scholar 

  14. Margulies, M. et al. Nature 437, 376–380 (2005).

    Article  CAS  Google Scholar 

  15. Howell, W.M., Jobs, M., Gyllensten, U. & Brookes, A.J. Nat. Biotechnol. 17, 87–88 (1999).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This study was supported by the EU Sixth Framework Programme integrated project MOLTOOLS (to A.J.B.).

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Authors and Affiliations

Authors

Contributions

L.S.M. and A.J.B. designed and planned the study. L.S.M. performed the majority of the laboratory experiments. J.P.P. produced the GS20 sequences. G.K., O.L. and A.J.B. performed data analysis of the GS20 data. L.S.M. and A.J.B. prepared the manuscript.

Note: Supplementary information is available on the Nature Methods website.

Corresponding author

Correspondence to Anthony J Brookes.

Ethics declarations

Competing interests

A.J.B. and the University of Leicester have filed a patent application covering the technology described in this article.

J.P.P. and G.K. are employed by Roche Applied Science.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–3, Supplementary Table 2, Supplementary Methods. (PDF 737 kb)

Supplementary Table 1

Oligonucleotide sequences. (XLS 90 kb)

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Meuzelaar, L., Lancaster, O., Pasche, J. et al. MegaPlex PCR: a strategy for multiplex amplification. Nat Methods 4, 835–837 (2007). https://doi.org/10.1038/nmeth1091

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