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    Pure Culture Preparation and Plating Technique PDF

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    Aman Nasir
    This document discusses techniques for preparing and maintaining pure cultures of microorganisms, including: 1) Spread plating, streak plating, and pour plating techniques for isolating pure cultures on agar plates from mixed samples. 2) Instruments like pipettes, inoculating needles, and loops are used to aseptically transfer microbes between media. 3) Short term cultures can be stored on slants in refrigeration for 1-3 months, while long term preservation requires freeze drying (lyophilization) of stock cultures.

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    Pure Culture Preparation and Plating Technique PDF

    Uploaded by

    Aman Nasir
    151 views17 pages
    This document discusses techniques for preparing and maintaining pure cultures of microorganisms, including: 1) Spread plating, streak plating, and pour plating techniques for isolating pure cultures on agar plates from mixed samples. 2) Instruments like pipettes, inoculating needles, and loops are used to aseptically transfer microbes between media. 3) Short term cultures can be stored on slants in refrigeration for 1-3 months, while long term preservation requires freeze drying (lyophilization) of stock cultures.

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    This document discusses techniques for preparing and maintaining pure cultures of microorganisms, including: 1) Spread plating, streak plating, and pour plating techniques for isolating pure cultures on agar plates from mixed samples. 2) Instruments like pipettes, inoculating needles, and loops are used to aseptically transfer microbes between media. 3) Short term cultures can be stored on slants in refrigeration for 1-3 months, while long term preservation requires freeze drying (lyophilization) of stock cultures.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    151 views17 pages

    Pure Culture Preparation and Plating Technique PDF

    Uploaded by

    Aman Nasir
    This document discusses techniques for preparing and maintaining pure cultures of microorganisms, including: 1) Spread plating, streak plating, and pour plating techniques for isolating pure cultures on agar plates from mixed samples. 2) Instruments like pipettes, inoculating needles, and loops are used to aseptically transfer microbes between media. 3) Short term cultures can be stored on slants in refrigeration for 1-3 months, while long term preservation requires freeze drying (lyophilization) of stock cultures.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 17

    Pure Culture Preparation and

    Plating Technique
    Introduction

    Culture mixed sample Isolated colony Pure culture


    Spreading technique Subculture
    Streaking technique
    Pouring technique
    Introduction
    • Pure culture: represents the growth of a
    single species of microorganism.
    • Subculture: transferring of microorganism
    from one culture medium to another by
    subculturing, using specific procedures and
    aseptic technique.
    • Asepsis: means free from a toxic condition
    resulting from the presence of
    microorganisms.
    Culture Transfer Instruments

    • A pipette is an instrument often used to


    transfer aliquots of culture, to prepare serial
    dilutions of microorganisms, and to dispense
    chemical reagents.
    • Inoculating needles and loops are used to
    aseptically transfer microorganisms from
    broth, slant, or agar cultures to other media.
    Maintenance of pure culture
    • Short-term maintenance:
    - generally between one to three months.
    - can often be achieved by storing slant cultures
    in the refrigerator at 4° to 10°C.
    • Long-term maintenance:
    - The best way to preserve many stock cultures
    for long periods is through lyophilization
    (freeze-drying).
    Plating Techniques
    Spread-Plate Technique
    1. A small volume of dilute bacterial mixture
    containing 100 to 200 cells or less is
    transferred to the center of an agar plate.
    2. spread evenly over the surface with a sterile,
    L-shaped glass rod.
    • The glass rod is normally sterilized by dipping
    in alcohol and flamed to burn off the alcohol.
    Spread-Plate Technique

    1 2
    Spread-Plate Technique
    Plating Techniques
    Streak- Plate Technique
    1. In this technique, the bacterial mixture is
    transferred to the edge of an agar plate with
    an inoculating loop and then streaked out
    over the surface in one of several patterns.
    2. The key principle of this method is that by
    streaking, a dilution gradient is established
    on the surface of the plate as cells are
    deposited on the agar surface.
    Streak- Plate Technique
    Plating Techniques
    Pour-Plate Technique
    1. The original sample is diluted several times to
    reduce the microbial population sufficiently
    to obtain separate colonies upon plating.
    2. The small volumes of several diluted samples
    are added to sterile petri plates and mixed
    with liquid media agar that has been cooled
    to about 48° to 50°C.
    3. used with bacteria and fungi.
    Pour-Plate Technique
    Pour-Plate Technique
    An inoculated plate is always incubated in an
    inverted position, Why????
    Procedure for Isolation of Pure
    Cultures

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