An automated method for the determination of deoxyribonuclease activity as exemplified by fractionation of the components of the medicament Varidase(R)
1993

Automated Method for Measuring Deoxyribonuclease Activity

publication Evidence: moderate

Author Information

Author(s): I. C. Locke, M. P. Ramsey, S. S. HilFf, B. G. Carpenter

Primary Institution: School of Pharmacy and Biomedical Sciences, University of Portsmouth

Hypothesis

Can an automated method effectively determine deoxyribonuclease activity despite the presence of inorganic cations?

Conclusion

The automated assay provides a more accurate profile of deoxyribonuclease activity by removing the inhibitory effects of sodium chloride.

Supporting Evidence

  • The automated method can overcome issues caused by inorganic cations.
  • Comparison with manual methods showed that the automated system provides a more realistic profile of nuclease activity.
  • Dialysis units were used to minimize the effects of sodium chloride on enzyme activity.

Takeaway

This study shows a new way to automatically measure how well certain enzymes break down DNA, which helps in understanding their activity better.

Methodology

The study used an automated Technicon AutoAnalyser to measure absorbance changes at 260 nm to determine nuclease activity from chromatographic fractions.

Limitations

The assay may not be suitable for restriction endonucleases that cut DNA at specific sites, as they produce minimal absorbance changes.

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