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This experiment demonstrated techniques for preparing a solid nutrient agar culture medium. Agar powder was weighed and dissolved in water before being sterilized in an autoclave. The resulting clear, opaque agar medium provided a firm surface for discrete bacterial colony growth. Proper sterilization was shown to be crucial for preventing microbial contamination of the culture medium.

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1K views

Lab Report

This experiment demonstrated techniques for preparing a solid nutrient agar culture medium. Agar powder was weighed and dissolved in water before being sterilized in an autoclave. The resulting clear, opaque agar medium provided a firm surface for discrete bacterial colony growth. Proper sterilization was shown to be crucial for preventing microbial contamination of the culture medium.

Uploaded by

Zemiah Puerto
Copyright
© © All Rights Reserved
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
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MICROBIAL ARTIFICIAL ENVIRONMENT

ABSTRACT

The purpose of this experiment was to demonstrate techniques for solid medium preparation using nutrient agar.
Several techniques was conducted in the agar medium preparation such as weighing of agar powder, liquid calibration,
dissolution of agar powder and sterilization using the autoclave. It was observed that the concentration and physical
structure of the solid medium agar makes it advantageous for the bacteria to discretely grow. The agar medium turned to
be clear opaque in color which implied that the medium was sterile. This result clearly supports that proper sterilization
was crucial to keep the culture media free from microbial contamination. Prolonged sterilization of discarded spent media
was also conducted to prevent the spread of harmful organisms.

INTRODUCTION completely solidified. Turn the petri dishes upside down


to prevent moisture.
The growth of microbial cells require very RESULTS AND DISCUSSION
specific environmental conditions, food, energy, proper
temperature and humidity. When culturing cells in the Liquid media are water-based solutions that do
laboratory, these conditions must be created using not solidify at temperatures above freezing and that tend
culture medium. A culture medium is a liquid or to flow freely when the container is tilted. Semisolid
gelatinous substance that contains essential nutrients to media exhibit a clotlike consistency because they
cultivate target microorganisms. contain an amount of solidifying agent that thickens them
Culture media can be classified based on their but does not produce a firm substrate. The result of this
composition as Synthetic and Non-synthetic medium. lab experiment shows that an agar medium is a perfect
Synthetic media contain pure chemical nutrients that example of Solid media which provide a firm surface on
vary little from one source to another and have a which cells can form discrete colonies and are beneficial
molecular content specified by means of an exact for culturing bacteria and fungi. It was observed that the
formula while the composition of Non-synthetic medium sterilization of the culture media in the autoclave
cannot be described by an exact chemical formula. One removed unwanted microorganisms that may
of the most commonly used culture media is the agar contaminate the solution.
medium. The aim of this experiment is to demonstrate
techniques for solid medium preparation using nutrient CONCLUSION AND RECOMMENDATION
agar.
Agar culture media can be prepared by following
MATERIALS AND METHODS the correct procedures. To test the sterility of media, no
growth of microbes should be observed. Hence, it is
First, add 100 ml of deionized water to a crucial to sterilize the media with proper care. When
graduated cylinder. Place 5.2g of agar on a weigh boat discarding spent media, it is necessary to prolong
or a weighing paper. Weigh out the agar powder using sterilization to prevent contamination of harmful bacteria.
an analytical balance. Now, place 75 ml of deionized Other ways of sterilization technique such as use of
water to a flask. Place the flask onto a stir plate then microwave oven and adding of antibacterial agents may
slowly pour the powder with rapid stirring to obtain a also be considered. However, appropriate consideration
uniform suspension. After about two minutes, add the needs to be given when using these techniques.
remaining 25 ml of water from the graduated cylinder.
Stir the medium until the agar powder is completely REFERENCES
dissolved or when it turns light amber in color. Next,
cover the flask with aluminum foil. Place a piece of Bassiri, E., Preparation of Media. Philadelphia, PN:
autoclave tape over the foil and label it with the media University of Pennsylvania, from
name.
https://www.sas.upenn.edu/LabManuals/biol275/Table
In the final step, use an autoclave to sterilize the
media. Power on the autoclave and ensure that drain _of_Contents_files/2-PreparationOfMedia.pdf
valve is closed. Add deionized water to the level Rao, S. Bacterial culture media. Karanataka, India: JJM
indicator line. Place the flask into the autoclave chamber Medical College, Department of Microbiology
then close the lid and turn the handle until it is secured https://www.microrao.com/micronotes/culture_media.pdf
tightly. Sterilize the medium at 121 °C for 15 minutes. Talaro, K.T. & Barry, C. (2015). Foundations in
After sterilizing, use heatproof gloves to remove the Microbiology (9th ed.) 2 Penn Plaza, New York, NY
flask. Dispense the medium into the petri dishes then 10121: McGraw-Hill Education
allow the medium to cool for up to 2 hours or until it is

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