PREPARATION OF MEDIA

Ms.A.Jeevarathinam M.Sc.,M.Phil.,P.G.D.C.A.,

Assistant Professor,

Department of Food Processing and Quality Control,

V.V.Vannia Perumal College for women,

Virudhunagar.

Aim

1. To become familiar with media preparation.

2. To cultivate microorganisms.

Introduction

A culture may be defined as growth of microbes under contamination free condition in

the laboratory on a suitable medium. Any materials that support the growth of organisms are
called culture media. It must contain many nutrients required for the growth of microorganisms.
The culture medium categorized into two depending on the consistency ie., solid or liquid..

Solid media can be prepared by the addition of agar to the medium which soildies at
room temperature. The solid surface of the medium is ideal for spores or cells of microbes as
they get trapped and grow very well.

Liquid media is also known as broth culture. A broth culture is defined as a sterile liquid
nutrient solution which does not have any solidifying agent.

Materials required

distilled water
conical flasks
cotton,
agar agar
pH meter
 test tube
glass rod
petriplates,
hot air oven
autoclave
Procedure

Nutrient broth

Nutrient broth is a general- purpose medium. This liquid medium is widely used for
general cultivation of aerobic bacteria and as a basal medium for a variety of physiological tests.

Composition (gm/litre)

Peptone 5
Yeast extract 1.5
Beef extract 1.5
Sodium chloride 5
Final pH 7.4 ± 0.2

13.0 grams of nutrient broth is suspended in 1000 ml of distilled water. The medium is
heated to promote solubilization of the ingredients. The medium is then sterilized by autoclaving
at 15 Lbs pressure for 15 minutes.

Nutrient agar

Nutrient agar is a general purpose solid medium. It is commonly used for isolation and
enumeration of bacteria from various substrates.

Composition (gm/litre)

Peptone 5
Yeast extract 1.5
Beef extract 1.5
Sodium chloride 5
Final pH 7.4 ± 0.2
Agar-Agar 15g

28.0 grams of nutrient agar is suspended in 1000ml of distilled water. The medium is
heated to promote solubilization of the ingredients. The medium is then sterilized by autoclaving
at 15 lbs pressure for 15minutes.