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Amplification of complex gene libraries by emulsion PCR

Nature Methods volume 3, pages 545–550 (2006)Cite this article

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  • ABIL EM 90, a surfactant (Degussa)

  • Bovine serum albumin (BSA) (for molecular biology, powder; Sigma-Aldrich)

  • Deoxynucleoside triphosphate (dNTP) mix (Roche Diagnostics)

  • Diethyl ether (water-saturated; ≥99.5%; Riedel-de Haën)

  • Ethyl acetate (water-saturated; ≥99.7%; Riedel-de Haën)

  • Mineral oil (for molecular biology, light oil; Sigma-Aldrich)

  • Pfu Turbo DNA polymerase (2.5 U/μl; Stratagene)

  • Span 80, a surfactant (Fluka)

  • Triton X-100, a surfactant (general-purpose grade; Fisher Scientific)

  • Tween 80, a surfactant (Sigma-Aldrich)

ABIL EM 90, a surfactant (Degussa)

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Figure 1: Amplification of complex gene libraries by conventional PCR and emulsion PCR.
Figure 2: Ethidium bromide–stained agarose gels reveal the absence of size competition and chimeric product–formation in ABIL EM 90–based emulsion PCRs.
Figure 3: Amplification of genomic DNA libraries by conventional PCR and emulsion PCR.

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Authors and Affiliations

  1. MRC Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 2QH, UK

    Richard Williams, Oliver J Miller & Andrew D Griffiths

  2. Department of Biological Chemistry, Weizmann Institute of Science, Rehovot, 76100, Israel

    Sergio G Peisajovich & Dan S Tawfik

  3. Institut de Science et d'Ingénierie Supramoléculaires (ISIS), 8 allée Gaspard Monge, BP 70028, Strasbourg Cedex, 67083, France

    Oliver J Miller & Andrew D Griffiths

  4. Casali Institute of Applied Chemistry, The Hebrew University of Jerusalem, Givat Ram, 91904, Jerusalem, Israel

    Shlomo Magdassi

Authors
  1. Richard Williams
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  5. Dan S Tawfik
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Corresponding authors

Correspondence to Dan S Tawfik or Andrew D Griffiths.

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Competing interests

The Medical Research Council (MRC) and the Weizmann Institute have filed patent applications, which include some of the ideas described in this manuscript. Should these organizations receive revenues from licensing these patents, the authors may be entitled to receive payments through the organizations’ Rewards to Inventors schemes.

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Williams, R., Peisajovich, S., Miller, O. et al. Amplification of complex gene libraries by emulsion PCR. Nat Methods 3, 545–550 (2006). https://doi.org/10.1038/nmeth896

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