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Uracil-DNA Glycosylase Assay by Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry Analysis
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1Department of Clinical Laboratory Sciences and Medical Biotechnology, College of Medicine,National Taiwan University, 2Department of Laboratory Medicine,National Taiwan University Hospital, 3Center for Microbial Pathogenesis, Nationwide Children’s Hospital and the Department of Pediatrics,The Ohio State University
Chapters
- 00:04Introduction
- 01:28DNA Glycosylase Assay
- 04:43Transferring UDG Reaction Products to a Matrix Chip
- 05:33Setting up the Assay Parameters on the Mass Spectrometer
- 07:19Mass Spectrometer Operation, Viewing Mass Spectra, and Analyzing the Data
- 09:03Results: UDG Assay Using MALDI-TOF MS for AP Site Containing Product Analysis
- 11:01Conclusion
A non-labeled, non-radio-isotopic method to assay uracil-DNA glycosylase activity was developed using MALDI-TOF mass spectrometry for direct apurinic/apyrimidinic site-containing product analysis. The assay proved to be quite simple, specific, rapid, and easy to use for DNA glycosylase measurement.
