| Chapter title |
Methylation-Specific PCR
|
|---|---|
| Chapter number | 23 |
| Book title |
DNA Methylation Protocols
|
| Published in |
Methods in molecular biology, January 2018
|
| DOI | 10.1007/978-1-4939-7481-8_23 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7479-5, 978-1-4939-7481-8
|
| Authors |
João Ramalho-Carvalho, Rui Henrique, Carmen Jerónimo |
| Abstract |
Cytosine methylation is a DNA modification generally associated with transcriptional silencing. Factors that regulate methylation have been linked to human disease, yet how they contribute to malignancies remains largely unknown. Methylation of DNA can change the functional state of regulatory regions, but does not change the Watson-Crick base pairing of cytosine. Moreover, sequence symmetry of CpGs enables propagation of the methyl mark through cell division. This potential for inheritance coupled with the fact that DNA methylation patterns change during development and disease partially explains the interest in DNA methylation as a memory module. DNA methylation analysis also provides an opportunity to discover new and more powerful biomarkers that can help in clinical practice.Methylation-Specific PCR (MSP) is likely the most widely used technique to study DNA methylation of a locus of interest. MSP can rapidly detect the methylation status of any group of CpG sites within a CpG island, not requiring methylation-sensitive restriction enzymes. It also requires minute amounts of DNA, is very sensitive as it can detect <0.1% of methylated alleles in a specific locus, and can be used in different samples, including bodily fluids, and paraffin-embedded samples. |
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