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    Laboratory Mediadispensing

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    leighzy lala
    The document describes the procedure for preparing microbiological media. It discusses that different media are used to culture different microorganisms and must be sterilized before use. The procedure involves adding ingredients like agar powder and nutrients to water in an Erlenmeyer flask. The mixture is then heated to dissolve the agar, poured into tubes, and autoclaved for sterilization before use. Proper preparation of media and sterile technique are important to avoid contamination.

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    Laboratory Mediadispensing

    Uploaded by

    leighzy lala
    41 views4 pages
    The document describes the procedure for preparing microbiological media. It discusses that different media are used to culture different microorganisms and must be sterilized before use. The procedure involves adding ingredients like agar powder and nutrients to water in an Erlenmeyer flask. The mixture is then heated to dissolve the agar, poured into tubes, and autoclaved for sterilization before use. Proper preparation of media and sterile technique are important to avoid contamination.

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    LABORATORY MEDIADISPENSING

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    The document describes the procedure for preparing microbiological media. It discusses that different media are used to culture different microorganisms and must be sterilized before use. The procedure involves adding ingredients like agar powder and nutrients to water in an Erlenmeyer flask. The mixture is then heated to dissolve the agar, poured into tubes, and autoclaved for sterilization before use. Proper preparation of media and sterile technique are important to avoid contamination.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    41 views4 pages

    Laboratory Mediadispensing

    Uploaded by

    leighzy lala
    The document describes the procedure for preparing microbiological media. It discusses that different media are used to culture different microorganisms and must be sterilized before use. The procedure involves adding ingredients like agar powder and nutrients to water in an Erlenmeyer flask. The mixture is then heated to dissolve the agar, poured into tubes, and autoclaved for sterilization before use. Proper preparation of media and sterile technique are important to avoid contamination.

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    © All Rights Reserved
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    of 4

    Activity No.

    2 Report Sheet

    MEDIA PREPARATION
    Name: Buenaflor, Andrea Mae P.
    Enorio, Isabelle D.
    Reyes, Shylene Kaye A.
    Rivera, NJ Aleah F.
    Yap, Fhardina D.

    I.Introduction
    Several kinds of microbiological media are used to cultivate bacteria and fungi. Depending
    on the microorganism that is being isolated or identified, a specific culture medium must be
    utilized. The medium may be given additional nutrients to increase its protein or sugar content.
    Several pH indicators are frequently used to differentiate bacteria depending on their metabolic
    activities; the indicators may change color when mildly basic or acidic. The medium may also be
    supplemented with growth stimulants, NaCl, and pH buffers to prevent it from deviating too far
    from neutral as the microorganisms metabolize.
    The traditional process for creating media involved adding each ingredient gradually, like
    in a recipe. Only when creating a customized medium to grow a particular picky organism where
    certain growth agents, nutrients, vitamins, and so forth have to be given in specific amounts.
    Chemically defined media is the name given to this medium (synthetic). And the most common
    bacteria that we want to cultivate can do nicely with the media that we regularly use in the lab.
    While some of our media is purchased.

    Nowadays, creating complicated media is actually very easy:


    • rehydrate the medium's powdered form
    • dissolve the agar powder, stir and boil the agar medium (if making an agar medium rather
    than a broth medium)
    • dispense the substance into tubes.
    • sterilize the tube medium with an autoclave.
    • pour the autoclaved agar medium for making plates into sterile petri dishes.

    Even after making microbiological media, it must be sterilized due to microbial


    contamination from air, glassware, hands, and other sources. Thousands of bacteria will reproduce
    in the medium within a few hours, so it must be sterilized quickly before the microbes start
    depleting the nutrients. The sterilization process ensures that the medium will remain sterile unless
    exposed to contaminants via a less-than-adequate aseptic technique (air exposure).

    Media should always be stored in a cold, moist environment to prevent evaporation,


    preferably in screw-capped tubes or bottles. Therefore, it is not advised to keep sterile media for
    an extended period of time unless stability has been proven. Media tubes should be reheated right
    before use if they have been stored for any period of time. To release dissolved gasses, liquid
    media should be heated for a short time in a boiling water bath or in flowing steam and then
    immediately cooled in cold water without agitation soon before inoculation. All bacteria prefer a
    damp surface, thus agar tubes should be melted and then allowed to solidify.

    II. Objectives
    • To learn about the environmental and nutritional requirements for cultivating organisms
    cultured in a lab.
    • To comprehend the use of an autoclave in the decontamination or sterilization process.
    • To become familiar with the methods for creating the media required for cultivating
    microorganisms.

    III. Requirements:
    A. Materials: Erlenmeyer Flask, 10mL Graduated Cylinder, Petri Plate, Electronic Balance,
    Hot Plate, Stirring Rod, Test Tubes, Test Tube Racks, Beaker, Autoclave
    B. Chemicals: Distilled Water, Media Base Powder

    III. Procedure
    1. Start by adding 250 ml of distilled water to a 500ml or 1L flask to make the TSB (broth).
    Turn on the hot plate and insert the stir bar to gently disrupt the surface. 3.25 grams of the
    TSB powder should be added to the flask, and it should dissolve (will happen quickly). At
    this point, there is no need to use heat.
    2. Pipette out 5 ml of the green cap after the powder has been dissolved.
    3. With the remaining solution (about 100ml) still stirring, add 2 grams of agar powder.
    4. The next step will require you to apply heat to the mixture. The agar has a strong tendency
    to boil over when it reaches 100⁰C. Watch the flask at all times once you see steam coming
    off of it.
    5. Remove the mixture from the heated plate as soon as it begins to boil (paper towels around
    the flask neck). DON'T just turn off the heat and leave the flask alone. Turning off the heat
    won't stop the flask from boiling over since the metal plate retains a lot of heat.
    6. The agar dissolves as it is heated up and turns a clearer, darker tan. The remainder of the
    agar medium in the flask will be poured into one large flask to save it for the preparation
    of petri plates after it has been taken off the heat and pipetted into 5 tubes for slants.
    7. Put the remaining melted agar and all of the tubes that you have pipetted out in the plastic
    autoclave racks.

    IV. Conclusion
    We came to the conclusion that there are essential things that should and shouldn't be done
    when preparing the cultured medium, the agar. The agar must be dissolved before heating the
    mixture in the Erlenmeyer flask with the distilled water because if not, the agar will burn. Then,
    the mixtures were heated in the microwave to completely dissolve the agar rather than on a hot
    plate since we did not have enough time. But because the Erlenmeyer flask that our group used
    was too large to fit in the microwave, we were unable to complete this step. This phase of the
    procedure was noted by our group, as we presumed that the absence of this could contribute to the
    results. The mixture was then autoclaved for sterilization.
    In preparing the broth, we dissolved the TSB powder in a 1 L flask containing 250 ml of
    distilled water. Hence, this distilled water is important as a medium through which nutrients can
    be transferred. Realizations have been made that, in doing the procedures, microbial contamination
    could easily happen. With that, ensuring proper protective apparel is important, as well as in
    quickly sterilizing the medium. Many sources and factors from the surroundings could affect the
    results of the laboratory activity.
    Documentation
    V. References
    • Microbial Culture Media Preparation. (2023). Merck. Retrieved March 9, 2023,
    from https://www.sigmaaldrich.com/PH/en/applications/microbiological-
    testing/microbial-culture-media-preparation
    • Mettler-Toledo International Inc. all rights reserved. (2022, November 16).
    Culture Media Preparation. Mettler-Toledo International Inc. All Rights
    Reserved.
    https://www.mt.com/ph/en/home/applications/Laboratory_weighing/Culture-
    Media-Preparation.html
    • Al-Tameemi, H. (2018, July 12). Medical Microbiology Laboratory (culture
    media preparation). https://www.slideshare.net/HusseinAltameemi2/medical-
    microbiology-laboratory-culture-media-preparation
    • Aryal, S. (2022, August 10). Nutrient Agar: Composition, Preparation and Uses.
    Microbiology Info.com. https://microbiologyinfo.com/nutrient-agar-composition-
    preparation-and-
    uses/#:~:text=Water%20is%20essential%20for%20the,various%20nutrients%20c
    an%20be%20transported.

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