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  • Surgery (Austin & Northern Health)
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    A new approach to primer design for the control of PCR bias in methylation studies.

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    100
    Author
    Wojdacz, TK; Hansen, LL; Dobrovic, A
    Date
    2008-07-28
    Source Title
    BMC Research Notes
    Publisher
    Springer Science and Business Media LLC
    University of Melbourne Author/s
    Dobrovic, Alexander
    Affiliation
    Surgery (Austin & Northern Health)
    Metadata
    Show full item record
    Document Type
    Journal Article
    Citations
    Wojdacz, T. K., Hansen, L. L. & Dobrovic, A. (2008). A new approach to primer design for the control of PCR bias in methylation studies.. BMC Res Notes, 1 (1), pp.54-. https://doi.org/10.1186/1756-0500-1-54.
    Access Status
    Open Access
    URI
    http://hdl.handle.net/11343/257291
    DOI
    10.1186/1756-0500-1-54
    Open Access at PMC
    http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2525644
    Abstract
    Primer design for PCR-based methylation analysis following bisulfite conversion of DNA is considerably more complex than primer design for regular PCR. The choice of the optimal primer set is critical to the performance and correct interpretation of the results. Most methodologies in methylation analysis utilize primers that theoretically amplify methylated and unmethylated templates at the same time. The proportional amplification of all templates is critical but difficult to achieve due to PCR bias favouring the amplification of the unmethylated template. The focus of this brief communication is to point out the important criteria needed for the successful choice of primers that will enable the control of PCR bias in bisulfite based methylation-screening protocols.

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