Abstract
Methylation at the C5 position of cytosine in DNA constitutes a prominent epigenetic modification. Reactive oxygen species (ROS) are ubiquitous within cells and they are very reactive toward 5-methyl-2′-deoxycytidine (5-mdC). Additionally, previous studies showed that 5-mdC could also be oxidized by ten-eleven translocation (Tet) family dioxygenases. Quantitative measurement of oxidation products of 5-mdC can facilitate the investigation about the roles of these modifications in gene regulation. Here we describe an LC-MS/MS/MS-based method for the sensitive and accurate quantifications of 5-hydroxymethyl-2′-deoxycytidine (5-HmdC), 5-formyl-2′-deoxycytidine (5-FodC), 5-carboxyl-2′-deoxycytidine (5-CadC), and 5-hydroxymethyl-2′-deoxyuridine (5-HmdU) in genomic DNA.
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Acknowledgement
The author thanks for the financial support from National Institutes of Health (NIH): NIH [CA210072].
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Yin, J., Liu, S., Wang, Y. (2022). Quantitative Assessment of the Oxidation Products of 5-Methylcytosine in DNA by Liquid Chromatography-Tandem Mass Spectrometry. In: Yuan, BF. (eds) DNA Modification Detection Methods . Springer Protocols Handbooks. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1229-3_1
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DOI: https://doi.org/10.1007/978-1-0716-1229-3_1
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