A new approach to primer design for the control of PCR bias in methylation studies.

BMC Research Notes
Tomasz K WojdaczAlexander Dobrovic

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.

References

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Aug 20, 2008·BMC Research Notes·Tomasz K WojdaczAlexander Dobrovic

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Citations

Jun 22, 2010·Breast Cancer Research and Treatment·Katie T HuangStephen B Fox
Jan 31, 2009·Nature Protocols·Tomasz K WojdaczLise Lotte Hansen
Dec 11, 2013·Nature Reviews. Genetics·Kimberly RobaskyGeorge M Church
Apr 14, 2011·Nucleic Acids Research·Evgeny A MoskalevJörg D Hoheisel
Dec 1, 2009·Epigenomics·Tomasz K Wojdacz
Jul 16, 2010·Expert Review of Molecular Diagnostics·Tomasz K WojdaczLise Lotte Hansen
Sep 18, 2014·Genes·Thomas Mikeska, Jeffrey M Craig
Apr 11, 2013·Expert Review of Molecular Diagnostics·Surbhi JainYing-Hsiu Su
Oct 22, 2014·Breast Cancer Research : BCR·Jia-Min B PangStephen B Fox
Oct 5, 2010·Clinical Biochemistry·Stefanie StanzerNadia Dandachi
Sep 3, 2010·Clinica Chimica Acta; International Journal of Clinical Chemistry·Benedetta IzziKathleen Freson
Sep 24, 2015·BMC Cancer·Giulia BrisottoManuela Santarosa
Jun 26, 2015·Journal of Clinical Laboratory Analysis·Loo Keat WeiLyn R Griffiths
May 17, 2015·Analytical Biochemistry·Yuki KonishiHiroko Higashimoto
Aug 20, 2008·BMC Research Notes·Tomasz K WojdaczAlexander Dobrovic
Jan 26, 2012·Epigenetics : Official Journal of the DNA Methylation Society·Michelle NewmanRebecca J Ormsby
Jun 29, 2013·Epigenetics : Official Journal of the DNA Methylation Society·Rodrigo M GonzálezNorberto D Iusem
Apr 11, 2015·Clinical Epigenetics·Darren KorbieMatt Trau
Aug 3, 2013·Progress in Neuro-psychopharmacology & Biological Psychiatry·Katharina DomschkeJürgen Deckert
May 26, 2016·Clinical Epigenetics·Claudia KnotheJörn Lötsch

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Methods Mentioned

BETA
PCR

Software Mentioned

Oligonucleotide Properties Calculator
Amplify
Primer Express

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