論文

査読有り 筆頭著者 責任著者
2011年

Highly efficient PCR assay to discriminate allelic DNA methylation status using whole genome amplification

BMC Research Notes
  • Yoichi Yamada
  • ,
  • Takashi Ito

4
179
開始ページ
179
終了ページ
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1186/1756-0500-4-179

Background: We previously developed a simple method termed HpaII-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. Findings. We developed HpaII-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 g. 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. Conclusions: HM-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. © 2011 Yamada et al
licensee BioMed Central Ltd.

リンク情報
DOI
https://doi.org/10.1186/1756-0500-4-179
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/21663670
ID情報
  • DOI : 10.1186/1756-0500-4-179
  • ISSN : 1756-0500
  • PubMed ID : 21663670
  • SCOPUS ID : 79958146649

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