Aim: Preparation of nutrient agar and nutrient broth media for cultivation of microorganisms.

MATERIALS REQUIRED:-
Peptones, Beef extract, NaCl, Agar, distilled water, Electron balance, test tube, measuring cylinder, pH meter,
heater, autoclave.
Nutrient Agar Medium Composition: 100 ml distilled water
Peptone- 5g/l
Beef extract-3g/l
NaCl-5g/l
pH- 7.0 ±0.02

Nutrient Broth Medium Composition: 100ml distilled water

Peptone- 5g/l
Beef extract-3g/l
NaCl-5g/l
pH- 7.0 ±0.02

PROCEDURE:-
1.) Weigh all the components of media using electronic balance (peptones, beef extract and Nacl) and then
mix with 100ml distilled water in a glass beaker.
2.) Check pH of the solution and maintain the pH- 7.0 ±0.02 by adding acid or base.
3.) Add agar to the mixture and heat.
4.) Autoclave at 121 °C for 20 minutes.
5.) Cool the medium and pour in petri-dishes/ test tubes.
6.) For nutrient broth medium all the components are added to distilled water in the same way except agar-
agar powder.

Nutrient agar and nutrient broth media is ready for culture growth.
PRECAUTIONS:-
1.) Handle the electronic balance carefully.
2.) Handle the pH meter and other instruments of laboratory carefully.
3.) Proper handling of the chemical components should be done.