UNIVERSITI KUALA LUMPUR

MALAYSIAN INSTITUTE OF CHEMICAL & BIOENGINEERING

TECHNOLOGY
LABORATORY TECHNICAL REPORT
SUBMISSION FORM

To: DR SHAHRUL ZAMAN Code Subject: CFB20303

From: Student ID. No.:
NASUHA ZAHIRAH BINTI ZAHARUDDIN 55218117094
NUR DIYANAH BINTI ABD JALIN 55218117050
NURAIN SYAKIRAH SABREENA BINTI A.RAHMAN 55218117062
NURUL ASYIKIN BINTI ABDUL LATIF
55218117105

No. of Group: 2 Date of Experiment: 18/2/2019

Title of Experiment: PREPARATION OF CULTURE MEDIA AND ASEPTIC TECHNIQUE IN
MICROBIOLOGY
Received by: Date of Submission: 1/3/2019

Note: Late submission will not be accepted.

*To be filled by the marker*

VERY VERY
POOR GOOD EXCELLENT
CRITERIA POOR GOOD
2 3 5
1 4
1.0 ABSTRACT & OBJECTIVES (HALF PAGE 2 4 6 8 10
ONLY) (TOTAL: 10%)
1. State the summary to the experiment
conducted.
2. State the objectives of the experiment (point
form)

2.0 PROCEDURES (TOTAL: 5%) 1 2 3 4 5

1.Methodology is presented in suitable and
understandable flowchart.
3.0 RESULTS (TOTAL: 10%) 2 4 6 8 10
1.Data are presented as deemed suitable with
complete label and units in tables and/or graphs.
4.0 DISCUSSIONS (MAXIMUM 1 PAGE) (TOTAL: 3 6 9 12 15
15%)
1. Explanations of the referred tables and/or
graphs are presented after it.
2. Discuss on the findings and relations to the
theory and objective of experiment.
5.0 CONCLUSIONS (TOTAL: 5%) 1 2 3 4 5
1. Summary of the results to relate the findings or
results with the theory applicable to the
experiment.
6.0 REFERENCES (TOTAL: 5%) 1 2 3 4 5
1. Minimum of 4 references.

TOTAL MARKS
ABSTRACT AND OBJECTIVES

The objectives of this experiment are to ensure student to be able to prepare microbiological media on
their own after completing this laboratory and students will be exposed to media preparation and
aseptic technique involve media preparation. In microbiology field, aseptic technique is such a
fundamental and important skill that must be mastered by the students. Proper aseptic technique is
required to prevent contamination of cultures from foreign bacteria in the environment. In working
with microorganism, the method of transferring growing organisms from pure culture to a sterile
medium is important. There are two parts in this experiment, Part A and Part B. In Part A, nutrients
agar were used as a medium. For part B, two technique were used which are aseptic and non-aseptic
technique. The result for aseptic technique and non-aseptic technique is different. There is no
presence of microorganism growth for aseptic technique while there are many microorganisms
growth for non-aseptic technique. Hence, aseptic technique is very important and useful in order to
avoid any contamination happen.
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
A. 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.
RESULTS

Aseptic Technique Non aseptic technique

Test 1

Test 2

Test 3

Test 4
DISCUSSION

Nowadays, the preparation of culture media using aseptic technique is commonly use in
microbiology for food industry. Proper aseptic technique can prevents contamination of cultures from
foreign bacteria inherent in the environment and prevents microbes used in the laboratory from
accidentally being released into the environment or infecting people working in the laboratories
(“Aseptic Technique and the Transfer of Microorganisms Objectives : Principle :,” 2019). The
objectives of this experiment are to make sure each student is able to prepare microbiological media
on their own and exposed to aseptic technique involve during media preparation. In part A, firstly,
10g of nutrient agar which act as a medium was weighed and mixed with 500 ml of distilled water.
The mixture was stirred with magnetic bar in stirring hot plate in order for the nutrient to dissolve
completely. Precaution steps need to be taken to ensure the mixture not burn on bottom or boil over.
Then, the mixture will be auto clave at 1 psi pressure and 121℃ to ensure the medium sterilized.
In part B, there were two technique used in these experiments which were aseptic and non-
aseptic technique. Bacteria were everywhere, and some were good for us while others are harmful.
Sterile work place, instrument and sample must be preventing from any contact to non-sterile
surfaces. This is to ensure the sample was not contaminated. Theoretically, there were no presences of
microorganism growth in aseptic technique’s sample. Based on the result obtained, in Test 1, 2, 3 and
4 for aseptic technique, there were no presence of microorganism growth in the sample as in the
theory. It shows that the procedures for aseptic technique have been done properly in this experiment.
Next, for result in Test 1, 2, 3 and 4 for non aseptic technique, there were many growth of
microorganism in the sample. It is because the samples are already exposed to the microorganism
during the experiment and cause it to be contaminated. Hence, aseptic technique was very important
and useful in order to avoid any contamination happen.
CONCLUSION

As for the conclusion, this experiment was done to make sure each student was able to prepare
microbiological media on their own. This experiment was also conducted to exposed to aseptic
technique involve during media preparation. The nutrient agar which act as a medium was used in the
experiment. Then, there were two types of technique used in these experiments which were aseptic
and non-aseptic technique. Based on the result obtained, the medium in Test 1, 2, 3 and 4 for aseptic
technique, there were no presence of microorganism growth in the sample but for non-aseptic
technique, there were many growths of microorganism in the sample. These shows that, there were no
presences of microorganism growth in aseptic technique’s sample because of work place and
instrument had been sterile. Furthermore, sample must be preventing from any contact to non-sterile
surfaces to make sure contamination not occurred. Lastly, the objectives of this experiment had been
achieved.
REFERENCES

1. Aseptic Technique and the Transfer of Microorganisms Objectives: Principle: (2019), 2–3.
Retrieved from http://vlab.amrita.edu/?sub=3&brch=73&sim=212&cnt=1
2. Aseptic Technique and the Transfer of Microorganisms Objective : Principle : (2019)
Retrieved from http://vlab.amrita.edu/?sub=3&brch=73&sim=212&cnt=1
3. Aseptic Technique and the Transfer of Microorganisms (Theory) : Microbiology Virtual Lab I
: Biotechnology and Biomedical Engineering : Amrita Vishwa Vidyapeetham Virtual Lab
http://vlab.amrita.edu/?sub=3&brch=73&sim=212&cnt=1
4. Aseptic Technique: Uses, Benefits, and Complications
https://www.healthline.com/health/aseptic-technique
5. Aseptic techniques | Nuffield Foundation
http://www.nuffieldfoundation.org/practical-biology/aseptic-techniques