Methylation Analysis
The AGRF offers a rapid and accurate technique to analyse methylation patterns using various platforms. Methylation and related chromatin changes are important processes in the regulation of gene expression. The relevance of DNA methylation has been demonstrated in mammalian development, imprinting and X-chromosome inactivation, suppression of parasitic DNA, and various cancer types. The ability to detect and to quantify methylation is particularly important to the field of cancer diagnostics. Changes in the methylation status of DNA have the potential to serve as an early detection marker for malignancies.
The benefits of quantitative methylation analysis is illustrated below using the Sequenom platform as an example:
- Rapid, automated discovery of multiple methylated CpG positions in regions of 200-600 bp
- Simplified assay design; no extension primers needed
- An alternative to problematic RNA extractions for gene expression determination
- No need for cloning of PCR products
- Quantitative assessment of the degree of methylation
- Simple identification of hypomethylation and hypermethylation
- Detection of 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
- High precision, reproducibility and throughput on an established MassARRAY system
Researchers provide reference sequence (up to 600bp per assay) and extracted DNA. SNP discovery assays are designed by AGRF, PCR oligos obtained and then processed, following PCR, on a mass spectrometer. The mass spectrometer provides a high level of sensitivity with methylation sites reported to the researcher in the form of a Microsoft Excel spreadsheet.
Sequenom Methylation Analysis Product Preview Note (PDF)
illumina DNA Methylation Analysis Data Sheet (PDF)
Methylation Analysis: Sample Submission Requirements for the Sequenom
For sample submission requirements for Methylation Analysis using other AGRF platforms, please contact us.
Methylation Analysis Service Process using the Sequenom Platform
illumina's Infinium Human Methylation27 BeadChip Assay features the following:
- Single CpG site resolution for 27 578 CpG sites encompassing more than 14000 genes
- Multi-sample format allows for processing of 12 samples on a single BeadChip
- BeadStudio Software analysis of methylation data
- Targets the promoter regions of 110 miRNA genes
Methylation Analysis Service Process using the illumina Platform
The AGRF platforms that can be used for Methylation Analysis are:
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