A new approach to primer design for the control of PCR bias in methylation studies.
AuthorWojdacz, TK; Hansen, LL; Dobrovic, A
Source TitleBMC Research Notes
PublisherSpringer Science and Business Media LLC
University of Melbourne Author/sDobrovic, Alexander
AffiliationSurgery (Austin & Northern Health)
Document TypeJournal Article
CitationsWojdacz, 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 StatusOpen Access
Open Access at PMChttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC2525644
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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