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Appl Environ Microbiol. 1969 August; 18(2): 174-176
Copyright © 1969 American Society for Microbiology. All Rights Reserved.

Detection of Nuclease Activity in Semisolid and Broth Cultures¹

^a Food Research Institute and Department of Bacteriology, College of Agricultural and Life Sciences, The Univesity of Wisconsin, Madison, Wisconsin 53706

ABSTRACT

A method for the detection of deoxyribonuclease activity in semisolid agar cultures was investigated. When cultures were overlaid with an acridine orangedeoxyribonucleate-agar (ADA) mixture, incubated for 1 to 3 hr, and observed under ultraviolet light, clear halos developed around colonies that produced deoxyribonuclease. A variation of the method for use with broth cultures involved impregnation of filter-paper discs with a small portion of the culture and overlaying the discs with the ADA mixture. This alteration has the advantage that the test tube cultures are easily heat-treated prior to assay to determine the heat resistance of the enzyme.

² Present address: Department of Microbiology, Weber State College, Ogden, Utah 84403.

¹ Published with the approval of the Director of the Wisconsin Agricultural Experiment Station.