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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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
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.
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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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DOI: https://doi.org/10.1038/nmeth1091
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