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Uracil-DNA Glycosylase Assay by Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry Analysis
JoVE 杂志
生物化学
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JoVE 杂志 生物化学
Uracil-DNA Glycosylase Assay by Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry Analysis
DOI:

12:19 min

April 22, 2022

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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

Summary

自动翻译

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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