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Uracil-DNA Glycosylase Assay by Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry Analysis

Article DOI: 10.3791/63089-v • 12:19 min • April 22nd, 2022
April 22nd, 2022 •
*1, *1,2, 3, 2, 1,2, 1,2, 1,2, 1,2
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
* These authors contributed equally

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Summary
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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.

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Uracil-DNA Glycosylase Assay Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry Analysis DNA Repair Mechanisms Genome Integrity Enzyme Activity Assay AP Product Label-free Assay High Specificity Versatility Scalability Rapidity E.coli Uracil-DNA Glycosylase DNA Glycosylase Assays Laboratory Personnel Pipetting Skills Dilution Skills Freshly Prepared Reagents Low Salt Buffers Background Noise Hui-Lan Chang PhD Student Guanine-uracil Base-bearing Substrate Of T1 U+9 Duplex
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