2011年
Highly efficient PCR assay to discriminate allelic DNA methylation status using whole genome amplification
BMC Research Notes
- ,
- 巻
- 4
- 号
- 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.
licensee BioMed Central Ltd.
- ID情報
-
- DOI : 10.1186/1756-0500-4-179
- ISSN : 1756-0500
- PubMed ID : 21663670
- SCOPUS ID : 79958146649