PROCEDURE

Agar Preparation Method

A. Weighing the Dry Agar

100ml of beaker have been tare and the Mass of dehydrated agar & tare weight
weight are recorded. have been total up.

Dry reagent was added. Edge of the

beaker were gently tap with spatula.
dH2O was added into the beaker until
The reagent bottle was tilted and rolled
the desired volume which is 70mL and
and its neck are tapped to sprinkle in
have been stirred to ensure no clumps
the final amount until it reach it's
of powder.
desired weight. The amount of the
reagent was recorded.

B. Dissolving the Media

The agar was stirred with a magnetic

The agar was poured into a 100mL
bar & heated with a hot plate to boiling
bottle. The bottle was cap loosely and
(95-100°C) without burning the bottom
been labelled.
or over boiling
C. Autoclaving the Media

Loosely capped agar was placed

in the autoclave

The autoclave are set up for slow

exhaust. The agar have been
autoclaved at 1psi. pressure for
15 minutes at 121°C.

The agar was removed from the

autoclave, and it is cooled down
to 50-60°C

Pouring the Agar

Petri dishes have been The sides of the dishes

take out carefully from
the preserving bag for
later storage.
Plates are poured by
have been mark
using sterile technique
according to code lines
on a sterile field.
to indicate type of agar

The lip of the bottle Packs of sterile plates

The lip of bottle are was flamed, the agar are placed near the
flamed whenever the was poured into the desk, the cap have
lid is replaced. plate at least half full been removed and
or 3/4 full. holded with fingers

When plates was

Bottle was rinsed It was stored in a
solidified, it was
immediately after labeled plastic bag at
placed in a incubator
pouring into the last 4°C, and was pre-
with a 37°C for 48
plate. warm before using.
hours.
Procedure with Aseptic Technique

The table top of laminar

Bunser burner have been
flow cabinet have been
lighted up
wiped with alcohol

The apparatus was

The inoculating loop was arranged close to the
placed in the alcohol Bunsen burner for 10
solution. minutes before the
experiment started

Sterile tips have been

The agar have been poured
picked up by using a
aseptically and the lid of
pipette and 0.1ml of
the dish have been closed
distilled water was
quickly. The agar was
dispensed aseptically and
allowed to cool and
transfered into nutrient
hardened.
broth.

Wire loop was flamed until

red and been cooled down.
The loop was dipped into
Lid was closed quickly.
distilled water and streak
S-shaped on the surface of
the agar aseptically.

The agar have been

Growth of
incubated in an incubator
microorganismsin agar and
and nutrient broth was
nutrient broth have been
incubated in incubator
observed
shaker for 24hrs at 37°C
Procedure without aseptic technique (Non-Aseptic)

Molten agar have been

Apparatus have been
poured into the petri dish by
arranged on the table
allowing it to flow across
without sterilizing the table.
the dish.

The steriled tips have been

been picked up by using
Lid of petri dish was closed
pipette. 0.1ml of distilled
and agar was allowed to
water was dispensed and
hardened
transfered into nutrient
broth

Agar plates have been

incubated in incubator and
Lid of petri dish was closed
nutrient broth was
quickly.
incubated in incubator
shaker for 24hrs at 37°C

Agar and nutrient broth

have been observed for the
growth of microorganisms.