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. 2012 Aug 27;10:173. doi: 10.1186/1479-5876-10-173

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Copyright ©2012 Bihl et al.; licensee BioMed Central Ltd.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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A): Serial dilutions of 100% and 0% methylated DNA were combined and pyrosequencing used to analyse the measured and theoretical percentage of methylated DNA as a function of the number of PCR cycles. A strong correlation between the measured and theoretical percentage of methylation was observed, particularly for PCR cycle numbers of 35 or more. B) Influence on methylation background as a function of PCR cycle numbers. C) In contrast to pyrosequencing, only a poor correlation between the measured and theoretical percentage of methylation was observed using methylation-specific PCR (MSP) which also showed a considerable dependency on PCR cycle number. D) Comparison of measured and theoretical percentage of methylation for 40 and 50 PCR cycles obtained using pyrosequencing and MSP.