DNA Methylation

DNA Methylation Analysis Overview

DNA methylation is considered to be one of the most important epigenetic modifications involved in disease generation. Several investigators have reported increased DNA methylation in cancer, typically in the promoter regions. In humans, DNA methylation occurs at CpG sites that are often found near promoter regions. DNA methylation results in conversion of cytosine in the CpG islands to 5-methyl cytosine. The status of methylation in these regions can have a major effect on the expression of these genes.

Accurate DNA Methylation Analysis Service

The Genotyping Department of BioServe utilizes the Sequenom® MassARRAY® EpiTYPER system to analyze methylation patterns in human DNA. The MassARRAY® EpiTYPER system uses mass spectrometry to detect and quantitate the base-specific cleavage products generated by the MassCLEAVE reaction. The DNA methylation analysis starts with bisulphite treatment of the genomic DNA. PCR primers are designed to generate a fragment of 200-600 bp, and one of the primers consists of T7 promoter sequence. In the next step, in vitro RNA transcription is performed on the reverse strand followed by base specific cleavage using RNase A that cleaves at base U or C. Cleavage products are generated for both U and C in separate reactions and analyzed on MALDI-TOF MS. EpiTYPER software analyzes and quantitates each cleavage product.

Source: www.sequenom.com

Using the EpiTYPER system, BioServe's DNA Methylation Analysis Service provides:

• Rapid, automated analysis of multiple methylated CpG positions in regions of 200-600 bp
• Quantitative assessment of the degree of DNA methylation
• Sensitive detection of DNA methylation levels as low as 5% in sample mixtures
• Results may be obtained from various sample types including frozen tissue, mouth swabs, and paraffin embedded tissue.

For more information regarding our services, please call 301-470-3362 or contact us at info@bioserve.com.

For general information on DNA methylation analysis, the Sequenom® EpiTYPER DNA Methylation Analysis Application Note can be found at Sequenom's DNA Methylation Analysis page.