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ACTIVITY 5
Preparation of Culture Media
Prepared by Prof. Mark Joseph B. Cano

INTRODUCTION

Culture medium/media are semi-solid media used to support the growth of microorganisms.
These contained essential nutrition to allow growth of the organism. Its gel structure allows
absorption of nutrients and supports formation of microbial colony. In this experiment, improvised
culture media will be prepared. This does not contain the actual nutrition requirement of microbes
but is enough to support colony growth.

LEARNING OUTCOMES

At the end of the experiment the learner must be able to:

1) determine the methods of preparing culture media;

2) identify the basic ingredients of culture medium; and
3) demonstrate proper ways disinfecting the culture medium.

COURSE MATERIAL

Learning Intervention

This Laboratory Activity, including its assessments can be performed by the student either by any
or all, or combinations of following learning methods and materials:

a) Critical Reading – The activity is fitted to be performed by students through mental

visualization and thought experiment. The module itself is
sufficient for students who have no access on actual laboratory,
internet or other relevant instructional materials.
b) Virtual Experiment – Students who have internet access can further learn by visiting
the virtual laboratory sites about Culture Media, as follows:
- http://learn.chm.msu.edu/vibl/
- https://vlab.amrita.edu/?sub=3&brch=73&sim=212&cnt
=1
- https://www.scienceprofonline.org/vmc/vmc-lab/vmc-
laboratory-differential-selective-bacterial-growth-
media.html
c) Actual Experiment – The activity can be actually performed in the laboratory or home.
d) Audiovisual clips – The student can watch any audio-video clips about Culture
Media, but especially the following links below:
- https://www.youtube.com/watch?v=cneascR3OEc
- https://www.youtube.com/watch?v=fzk_-O2SDos

Materials:

• Clean Small Glass/plastic container with lid/ cover

• Agar powder (commercial gelatin powder; colorless/flavorless)

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• Potato (one piece)

• Distilled water (100ml)
• Bleach (zonrox)
• Gauze
• Candle
• Match sticks

Methodology:

Surface-Disinfection of small glass / plastic container

Disinfect glass/plastic container and lid by soaking them with bleach solution (1 cap bleach:
100mL water) for 10 minutes. Wash with distilled water the dry the container by placing them over
a clean tissue. Cover the container with lid without touching the inner part of the container and
bottom part of the lid. Ensure that hands are thoroughly washed with soap and water, or if
available, gloves are used while performing this part of the experiment. The container must remain
tightly closed until needed.

Preparation of Agar media

Wash, peel and cut the potato into halves o quarter depending on the size of the potato in 100mL
distilled water (use measuring cup or plastic bottles with indicated volume to measure water). Boil
the potato estimated for 15 minutes and remove after the potato becomes tender. To the potato
broth, add a teaspoon of agar powder, stir and wait until completely dissolves it by boiling. Add
small amount of distilled water if the volume decreases due to evaporation. Stir again and check
if the agar is somewhat sticky. Remove the undissolved agar by using the gauze or clean cloth
then cover the pot.

Pour-plating

First, sterilize the tabletop by spraying with alcohol then wipe the surface with clean cloth or tissue.
Next, put on some gloves and other protective barrier. Prepare the materials then placed in the
cleaned tabletop. Sterilize the sides if you are using a glass container by quickly passing over the
flamed candle in the circumference of the container/lid area. If in case you are using a plastic
container just place the candle in the radius close to your working area. Partially open the lid of
the container with lighted candle at the side when pouring the agar. Fill about half or ¾ of the
container then close. Occasionally, check if the media has successfully solidified. Do not open
the container upon checking. Just wiggle the container to determine if the agar already solidifies.
Invert the container once the agar has formed.

*If refrigerator is available, the media can be stored but must be placed in inverted position. Then,
the learner can proceed to the next experiment.

GENERAL MICROBIOLOGY laboratory

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ACTIVITY 5
PREPARATION OF CULTURE MEDIA

Name: ______________________________________ Score: ________________

Course, Year & Section: _______________________ Date: _________________

In a separate short white bond paper, write all the questions and answers with student
name, course, year and section, and title of the activity. Kindly write the questions given
then answer.

ACTIVITY / ASSESSMENT

1) Illustrate and describe the step by step method of preparing culture media
2) Search and explain for the significance of the following:
a. Why do we need to soak the container in a bleaching solution?
b. Why do we need to keep the container always closed except when pouring the
medium?
c. Why do we need to partially open the container during pour-plating?
d. What is the use of potato in the experiment?
e. Why do we need to Invert the container once the media/agar/gelatin had solidified?

GRADING SYSTEM

• Part 1 – Illustration. Accurately illustrate and describe the step-by-step process in

methodology. (2 points each)

Each item consists of 2 points each, a total maximum score creditable is 20 points.

• Part 2, which is a constructed-response assessment, the following rubric shall be used to

evaluate the answers in each items:

3 points 2 points 0 – 1 point

The answer is characterized The answer is characterized The answer is characterized
as: as: as:
• Having complete, correct • Having incomplete, yet • Incorrect and inaccurate;
idea/thought required by correct idea/thought • Irrelevant response; and
the question required by the question; • Unintelligible and cannot
• Concise and direct-to-the- • Not concise response be understand
point response • Grammar construction and
• Having syntax like somewhat obscures
construction and like which the response
doesn’t affect effective
communication

Each item consists of 3 points, thus the total maximum score creditable is 15 points.

Part 1 = 20 points
Part 2 = 15 points
Total = 35 points

GENERAL MICROBIOLOGY laboratory