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Continuous Fluorescence-Based Endonuclease-Coupled DNA Methylation Assay to Screen for DNA Methyltransferase Inhibitors

Article DOI: 10.3791/62949-v • 06:07 min • August 5th, 2022
August 5th, 2022 •
1, 2, 2
1Department of Chemistry, Bucknell University, 2Program in Cell Biology/Biochemistry, Bucknell University

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DNA methyltransferases are potential cancer drug targets. Here, a protocol is presented to assess small molecules for DNA methyltransferase inhibition. This assay utilizes an endonuclease to couple DNA methylation to fluorescence generation and allows for enzyme activity to be monitored in real time. 

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Fluorescence-based Assay Endonuclease-coupled Assay DNA Methylation Assay Screen For Inhibitors Methyltransferase Inhibitors Plate Reader Assay Continuous Real-time Data Collection Assay Conditions Methylation Buffer Bovine Serum Albumen (BSA) Hairpin DNA Substrate S-adenosylmethionine (SAM) Inhibitor Concentration Dimethyl Sulfoxide (DMSO) Control Sample Vortexing Mini Centrifuge 96-well Plate Gla I Enzyme
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