Application of 5-Methylcytosine DNA Glycosylase to the Quantitative Analysis of DNA Methylation
Abstract
:1. Introduction
2. Results
2.1. DME Induces Single-Strand DNA Breaks Proportional to DNA Methylation Density
2.2. DME-qPCR Allows a Quantitative DNA Methylation Analysis
2.3. DME-qPCR Distinguishes DNA Methylation Levels at the FWA Gene in Wild Type and Late Flowering Mutants in Arabidopsis
2.4. DME-qPCR Detects Changes in DNA Methylation Levels at the CNR Gene during Fruit Ripening in Tomato
3. Discussion
4. Materials and Methods
4.1. Protein Expression and Purification
4.2. DME Treatment and Bal-31 Digestion Followed by PCR on Methylated DNA Fragments
4.3. Comparison of DME Treatment and McrBC Digestion
4.4. Plant Materials
4.5. CAPS Analysis
4.6. Reverse Transcriptase-qPCR
4.7. Locus-Specific Bisulfite Sequencing
4.8. DME-qPCR
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Choi, W.L.; Mok, Y.G.; Huh, J.H. Application of 5-Methylcytosine DNA Glycosylase to the Quantitative Analysis of DNA Methylation. Int. J. Mol. Sci. 2021, 22, 1072. https://doi.org/10.3390/ijms22031072
Choi WL, Mok YG, Huh JH. Application of 5-Methylcytosine DNA Glycosylase to the Quantitative Analysis of DNA Methylation. International Journal of Molecular Sciences. 2021; 22(3):1072. https://doi.org/10.3390/ijms22031072
Chicago/Turabian StyleChoi, Woo Lee, Young Geun Mok, and Jin Hoe Huh. 2021. "Application of 5-Methylcytosine DNA Glycosylase to the Quantitative Analysis of DNA Methylation" International Journal of Molecular Sciences 22, no. 3: 1072. https://doi.org/10.3390/ijms22031072