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Research Article

Optimized PCR Conditions and Increased shRNA Fold Representation Improve Reproducibility of Pooled shRNA Screens

  • Žaklina Strezoska,

    Affiliation: Molecular Biology, Thermo Fisher Scientific, Lafayette, Colorado, United States of America

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  • Abel Licon,

    Affiliation: Molecular Biology, Thermo Fisher Scientific, Lafayette, Colorado, United States of America

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  • Josh Haimes,

    Affiliation: Molecular Biology, Thermo Fisher Scientific, Lafayette, Colorado, United States of America

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  • Katie Jansen Spayd,

    Affiliation: Molecular Biology, Thermo Fisher Scientific, Lafayette, Colorado, United States of America

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  • Kruti M. Patel,

    Affiliation: Molecular Biology, Thermo Fisher Scientific, Lafayette, Colorado, United States of America

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  • Kevin Sullivan,

    Affiliation: Molecular Biology, Thermo Fisher Scientific, Lafayette, Colorado, United States of America

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  • Katarzyna Jastrzebski,

    Affiliation: Victorian Centre for Functional Genomics, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia

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  • Kaylene J. Simpson,

    Affiliations: Victorian Centre for Functional Genomics, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia, The Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia

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  • Devin Leake,

    Affiliation: Molecular Biology, Thermo Fisher Scientific, Lafayette, Colorado, United States of America

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  • Anja van Brabant Smith,

    Affiliation: Molecular Biology, Thermo Fisher Scientific, Lafayette, Colorado, United States of America

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  • Annaleen Vermeulen mail

    annleen.vermeulen@thermofisher.com

    Affiliation: Molecular Biology, Thermo Fisher Scientific, Lafayette, Colorado, United States of America

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  • Published: August 01, 2012
  • DOI: 10.1371/journal.pone.0042341

About the Authors

Žaklina Strezoska, Abel Licon, Josh Haimes, Katie Jansen Spayd, Kruti M. Patel, Kevin Sullivan, Devin Leake, Anja van Brabant Smith, Annaleen Vermeulen
Molecular Biology, Thermo Fisher Scientific, Lafayette, Colorado, United States of America
Katarzyna Jastrzebski, Kaylene J. Simpson
Victorian Centre for Functional Genomics, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia
Kaylene J. Simpson
The Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia

Corresponding Author

Email: annleen.vermeulen@thermofisher.com

Competing Interests

Z. Strezoska, A. Licon, J. Haimes, K.J. Spayd, K.M. Patel, K. Sullivan, D. Leake and A. Vermeulen are employed or were formerly employed by Thermo Fisher Scientific, one of the funders of this study. Some of the materials used in this study are products sold by Thermo Fisher Scientific (including Decode RNAi Annotated Genome Screening Library, Nanodrop 1000, Phusion® Hot Start II High-Fidelity DNA Polymerase, ABsolute™ Blue qPCR SYBR® Green master mix and Matrix PlateMate) and are subject to use restrictions or licenses. There are no further patents, products in development or marketed products referenced in this article to declare. This does not alter the authors' adherence to all the PLoS ONE policies on sharing data and materials, as detailed online in the guide for authors. K. Jastrzebski and K.J. Simpson declare that they have no competing interests.

Author Contributions

Conceived and designed the experiments: ŽS AL JH K. J. Spayd KMP K. J. Simpson KJ DL AvBS AV. Performed the experiments: ŽS JH K. J. Spayd KMP. Analyzed the data: ŽS JH K. J. Spayd KMP KS AL DL AvBS AV. Contributed reagents/materials/analysis tools: AL KS. Wrote the paper: ZS JH K. J. Spayd KMP AL DL AvBS AV.