Quantitative measurement of genome-wide DNA methylation by a reliable and cost-efficient enzyme-linked immunosorbent assay technique
| Autor: | D. Kremer, S. Metzger, V. Kolb-Bachofen |
|---|---|
| Rok vydání: | 2012 |
| Předmět: |
Keratinocytes
Spectrometry Mass Electrospray Ionization Cost-Benefit Analysis Medizin Biophysics Enzyme-Linked Immunosorbent Assay Computational biology Biology Biochemistry Cytosine chemistry.chemical_compound Cell Line Tumor Humans Epigenetics Methylated DNA immunoprecipitation Molecular Biology Regulation of gene expression Genome Human DNA Cell Biology Methylation DNA Methylation Molecular biology Histone Gene Expression Regulation chemistry DNA methylation 5-Methylcytosine biology.protein Illumina Methylation Assay Biologie |
| Zdroj: | Analytical Biochemistry. 422:74-78 |
| ISSN: | 0003-2697 |
| DOI: | 10.1016/j.ab.2011.11.033 |
| Popis: | DNA methylation, the conversion of cytosine to 5-methylcytosine, is an important epigenetic modification involved in gene regulation. DNA methylation is essential for normal development whereas abnormal methylation has been implicated in pathological conditions including cancer. To evaluate the extent and variation of genome-wide DNA methylation and its changes during cellular differentiation and tumorgenesis as well as the interplay with histone modifications, accurate and reproducible quantification of the genomic DNA methylation level is required. These measurements have so far been achieved only by sophisticated and costly techniques. Here we report the generation of an enzyme-linked immunosorbent assay (methDNA-ELISA) for the accurate quantification of global DNA methylation levels. The linear region of this methDNA-ELISA ranges from 1 to 10%, making it highly suitable for the typical ranges from 2 to 6% in mammalian genomes. This method requires 10 ng of isolated DNA per sample, thus permitting investigation with minimal amounts of DNA previously not applicable for global DNA methylation analysis, e.g., clinical biopsies or cells collected by microdissection. |
| Databáze: | OpenAIRE |
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