DNAse Test

1. Principle of DNAse Test

DNA Hydrolysis test or Deoxyribonuclease (DNAse) test is used to determine the
ability of an organism to hydrolyze DNA and utilize it as a source of carbon and
energy for growth.

2. Indicator

An agar medium; DNase agar, a differential medium is used to test the ability of an
organism to produce deoxyribonuclease or DNase. This medium is pale green in
color because of DNA-methyl green (indicator) complex

If the organism that grows in the medium produces Deoxyribonuclease, it breaks

down DNA into smaller fragments.  When the DNA is broken down, it no longer
binds to the methyl green, and green color fades and the colony is surrounded by a
colorless zone

3. Result
A. Positive: When DNA is hydrolysed, methyl green is released turning the medium
colorless around the test organism.
B. Negative: If there is no degradation of DNA, the medium remains green.

4. Procedure and Requirement

Requirements:

1. Media: DNase Agar or DNase agar with Methyl green indicator.

2. Reagent: Hydrochloric acid (1mol/L) only when DNase agar without indicator is
used
3. Others: Inoculating loop, Bunsen burner
Procedure of DNase (DNA hydrolysis test)

1. Dry the surface of agar plates before use. Each plate may be divided into sections
by drawing lines on the bottom of the plate.
2. Inoculate the test agar medium: There are two types of inoculation that can be
done.

Spot Inoculation

 Touch a colony of the organism under test with a loop and inoculate it onto a
small area of the DNase test agar plate, in the middle of one of the marked
sections to form a thick plaque of growth 5-10 mm in diameter after incubation.
 Incubate the plate at 37°C for 18-24hr.

Band or line streak inoculation

 Use a heavy inoculum and draw a line 3-4 cm long from the rim to the centre of
the DNase test agar plate
 Incubate the plate at 37°C for 18-24hr.