Improved Quantification of DNA Methylation Using Methylation-Sensitive Restriction Enzymes and Real-Time PCR

  title={Improved Quantification of DNA Methylation Using Methylation-Sensitive Restriction Enzymes and Real-Time PCR},
  author={Ko Hashimoto and Shoichi Kokubun and Eiji Itoi and Helmtrud I. Roach},
  pages={86 - 91}
Heterogeneity of cells with respect to the DNA methylation status at a specific CpG site is a problem when assessing methylation status. We have developed a simple two-step method for the quantification of the percent of cells that display methylation at a specific CpG site in the promoter of a specific gene. The first step is overnight digestion of genomic DNA (optimal conc. 20ng/5μl) with a relevant methylation-sensitive restriction enzyme (optimal 2 units). This is followed by real time PCR… 
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Methylation-sensitive enrichment of minor DNA alleles using a double-strand DNA-specific nuclease
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A real-time PCR assay for DNA-methylation using methylation-specific blockers.
These assays show that HeavyMethyl technology can be successfully employed for the analysis of very low concentrations of methylated DNA, e.g. in serum of patients with tumors, and showed a highly significant methylation difference between normal colon and colon adenocarcinomas.
Combination of methylated-DNA precipitation and methylation-sensitive restriction enzymes (COMPARE-MS) for the rapid, sensitive and quantitative detection of DNA methylation
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Methylation-sensitive polymerase chain reaction.
Here, we describe a robust and reproducible methylation-sensitive polymerase chain reaction (MS-PCR) method to detect the percentage methylation in repeat sequences of individual pre-implantation
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Very little information is available on the methylation status of particular CpG sites in the promoter regions of specific genes from different cell types or cells from different developmental stages, due to several factors.
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