Methylation specific oligonucleotide microarray
Methylation specific oligonucleotide microarray (MSO microarray) is a high-throughput technique used to analyze DNA methylation patterns across the genome. This method combines the principles of microarray technology with methylation-specific detection, allowing researchers to study epigenetic modifications on a large scale.
Overview
DNA methylation is a crucial epigenetic modification involving the addition of a methyl group to the cytosine or adenine nucleotides in DNA. This process plays a significant role in regulating gene expression, genomic imprinting, X-chromosome inactivation, and suppression of transposable elements. Abnormal DNA methylation patterns are associated with various diseases, including cancer, neurological disorders, and cardiovascular diseases.
Principle
The MSO microarray technique involves several key steps: 1. DNA Extraction: Genomic DNA is extracted from the sample of interest. 2. Bisulfite Treatment: The DNA is treated with sodium bisulfite, which converts unmethylated cytosines to uracil, while methylated cytosines remain unchanged. 3. PCR Amplification: The bisulfite-treated DNA is amplified using polymerase chain reaction (PCR) with primers specific to methylated or unmethylated sequences. 4. Hybridization: The amplified DNA is hybridized to an oligonucleotide microarray containing probes specific to methylated and unmethylated sequences. 5. Detection and Analysis: The hybridized microarray is scanned, and the data is analyzed to determine the methylation status of the target sequences.
Applications
MSO microarrays are used in various research and clinical applications, including:
- Cancer Research: Identifying methylation biomarkers for early detection, prognosis, and therapeutic targets.
- Epigenetic Studies: Investigating the role of DNA methylation in gene regulation and development.
- Disease Diagnosis: Developing diagnostic tools for diseases associated with abnormal methylation patterns.
- Pharmacogenomics: Studying the impact of DNA methylation on drug response and personalized medicine.
Advantages and Limitations
Advantages
- High-throughput: Allows simultaneous analysis of thousands of CpG sites.
- Sensitivity: Capable of detecting low levels of methylation.
- Specificity: Differentiates between methylated and unmethylated sequences.
Limitations
- Cost: Relatively expensive compared to other methylation analysis methods.
- Complexity: Requires specialized equipment and expertise.
- Limited Coverage: May not cover all CpG sites in the genome.
See Also
References
External Links
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Contributors: Prab R. Tumpati, MD