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

3.7K Views

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12:19 min

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April 22nd, 2022

DOI :

10.3791/63089-v

April 22nd, 2022

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Hui-Lan Chang*¹, Kang-Yi Su*¹^,², Steven D. Goodman³, Wern-Cherng Cheng², Liang-In Lin¹^,², Ya-Chien Yang¹^,², Sui-Yuan Chang¹^,², Woei-horng Fang¹^,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

Summary

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Uracil DNA Glycosylase

DNA Repair

MALDI TOF Mass Spectrometry

Enzyme Activity Assay

AP Product

Label free Method

E coli Uracil DNA Glycosylase

Chapters in this video

0:04

Introduction

1:28

DNA Glycosylase Assay

4:43

Transferring UDG Reaction Products to a Matrix Chip

5:33

Setting up the Assay Parameters on the Mass Spectrometer

7:19

Mass Spectrometer Operation, Viewing Mass Spectra, and Analyzing the Data

9:03

Results: UDG Assay Using MALDI-TOF MS for AP Site Containing Product Analysis

11:01

Conclusion

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