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Appl Environ Microbiol. 1972 December; 24(6): 920-923
Copyright © 1972 American Society for Microbiology. All Rights Reserved.

Convenient Assay for Staphylococcal Nuclease by the Metachromatic Well-Agar-Diffusion Technique

Section of Infectious and Immunologic Diseases, Department of Internal Medicine, School of Medicine, University of California, Davis, California 95616
Department of Epidemiology and Preventive Medicine, School of Veterinary Medicine, University of California, Davis, California 95616

ABSTRACT

The metachromatic agar-diffusion (MAD) microslide technique was adapted for quantitative assay for staphylococcal thermonuclease in heterogeneous systems, such as milk and broth. When an enzyme-containing solution was placed in a well cut in the agar, a bright pink halo was obtained. The diameter of the pink zone of hydrolysis was related to time and temperature of incubation and to nuclease concentration. Concentrations of nuclease as low as 0.005 µg/ml and as high as 2.0 µg/ml were conveniently determined after 3 hr at 37 C.