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BMC Research Notes

, 4:179

First Online: 10 June 2011Received: 27 September 2010Accepted: 10 June 2011DOI: 10.1186-1756-0500-4-179

Cite this article as: Yamada, Y. & Ito, T. BMC Res Notes 2011 4: 179. doi:10.1186-1756-0500-4-179

Abstract

BackgroundWe previously developed a simple method termed Hpa II-McrBC PCR HM-PCR to discriminate allelic methylation status of the genomic sites of interest, and successfully applied it to a comprehensive analysis of CpG islands CGIs on human chromosome 21q. However, HM-PCR requires 200 ng of genomic DNA to examine one target site, thereby precluding its application to such samples that are limited in quantity.

FindingsWe developed Hpa II-McrBC whole-genome-amplification PCR HM-WGA-PCR that uses whole-genome-amplified DNA as the template. HM-WGA-PCR uses only 1-100th the genomic template material required for HM-PCR. Indeed, we successfully analyzed 147 CGIs by HM-WGA-PCR using only ~300 ng of DNA, whereas previous HM-PCR study had required ~30 μg. Furthermore, we confirmed that allelic methylation status revealed by HM-WGA-PCR is identical to that by HM-PCR in every case of the 147 CGIs tested, proving high consistency between the two methods.

ConclusionsHM-WGA-PCR would serve as a reliable alternative to HM-PCR in the analysis of allelic methylation status when the quantity of DNA available is limited.

Electronic supplementary materialThe online version of this article doi:10.1186-1756-0500-4-179 contains supplementary material, which is available to authorized users.

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Autor: Yoichi Yamada - Takashi Ito

Fuente: https://link.springer.com/







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