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Preserving Microbial
Cultures: Top 5
Methods
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The following points highlight the top five

methods of preserving microbial culture.
The methods are: 1. Agar Slant Cultures 2.
Agar Slant Culture Covered with Oil (Parafin
Method) 3. Saline Suspension 4.
Preservation at Very Low Temperature 5.
Preservation by Drying in Vacuum 6.
Preservation by Freeze Drying
(Byophilization).

Preserving Microbial Culture: Method

# 1.

Agar Slant Cultures:

All microbiology laboratories preserve

micro-organisms on agar slant. The slants
are incubated for 24hr or more and are then
stored in a refrigerator. These cultures are
periodically transferred to fresh media. Time
intervals at which the transfers are made
which varies with the origin and condition of
growth.
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Preserving Microbial Culture: Method
# 2.
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Agar Slant Culture Covered with Oil

(Parafin Method):

The agar slants are inoculated and incubated

until good growth appears. They are then
covered with sterile mineral oil to a depth of
1 cm above the tip of slant surface. Transfers
are made by removing a loop full of the
growth, touching the loop to the glass
surface to drain off excess oil, inoculating a
fresh medium and then preserving the initial
stock culture.

This is a simple and most economical

method of preserving bacteria and fungi
where they remain viable for several years at
room temperature. The layer of paraffin
prevents dehydration of the medium and by
ensuring an aerobic condition, the
microorganism remain in dormant state.

Preserving Microbial Culture: Method

# 3.

Saline Suspension:

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Sodium chloride in high concentration is

frequently an inhibitor of bacterial growth.
Bacteria are suspended in 1% salt solution
(sublethal concentration in screw cap tubes
to prevent evaporation). The tubes are
stored at room temperature. Whenever
needed the transfer is made on agar slant.

Preserving Microbial Culture: Method

# 4.

Preservation at Very Low

Temperature:

The organisms are suspended in nutrient

broth containing 15% glycerol. The
suspension is frozen and stored at -15°C to
-30°C. The availability of liquid nitrogen
(temp -196°C) provides another main
preserving stock culture. In this procedure
culture are frozen with a protective agent
(glycerol or dimethane sulphoxide) in sealed
ampoules. The frozen culture are kept in
liquid nitrogen refrigerator.

Preserving Microbial Culture: Method

# 5.

Preservation by Drying in Vacuum:

The organisms are dried over calcium

chloride in vacuum and are stored in the
refrigerator.

Preserving Microbial Culture: Method

# 6.
Preservation by Freeze Drying
(Byophilization):

In this process the microbial suspension is

placed in small vials. A thin film is frozen
over the inside surface of the vial by rotating
it in mixture of dry ice (solid carbon dioxide)
and alcohol, or acetone at a temperature of
−78oC .The vials are immediately connected
to a high vacuum line. This dries the
organism while still frozen. Finally, the
ampules are sealed off in a vacuum with
small flame.

These culture can be stored for several years

at 40°C. This method is also employed for
preservation of toxins, sera, enzymes and
other biological material. To revive
microbial cultures it is merely necessary to
break open the vial aseptically, add a
suitable stale medium, and after incubation
make further transfers.

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The process permits the maintenance of

longer number of culture without variation
in characteristics of the culture and greatly
reduces the danger of contamination.

Depending on the chemical constituents

from which they are made, their physical
nature and their functions, different
parameters of media are described here.
Related Articles:
1. Top 6 Methods Used for Preservation of
Organisms
2. Maintenance and Preservation of Pure
Cultures: 4 Methods

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TABLE OF CONTENTS

Preserving Microbial Culture:

Method # 1.
Preserving Microbial Culture:
Method # 2.
Preserving Microbial Culture:
Method # 3.
Preserving Microbial Culture:
Method # 4.
Preserving Microbial Culture:
Method # 5.
Preserving Microbial Culture:
Method # 6.

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