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    Lab 3B Isolation of Pure Cultures

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    Lab 3B Isolation of Pure Cultures

    Uploaded by

    brucehung
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    © © All Rights Reserved

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    Copyright:

    © All Rights Reserved
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    Download as pptx, pdf, or txt
    10 views7 pages

    Lab 3B Isolation of Pure Cultures

    Uploaded by

    brucehung

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    of 7

    Fall 2020 Microbiology

    Lab 3B
    Exp 3 Isolation of Pure Culture

    Streak plate for isolation.

    Fall 2020 Microbiology Lab #3


    Exp 3 Techniques for Isolation of
    Pure Culture

    Pure Culture – contains only one type of


    microbes.

    Streak Plates for Isolation

    • Use an inoculating loop to streak a mixed


    culture on a plate and spread the microbes on
    the plate.
    • The goal is to deposit single cells on the plate
    so that each single cell will divide and form a
    pure bacterial colony.
    Fall 2020 Microbiology Lab #3
    Demonstration Video

    Aseptic Technique: Inoculating a Petri Plate - Streaking for Iso


    lation
    . (5:58)

    Dr. Gary Kaiser demonstrates a T-streak technique to spread bacteria on a


    plate to get single cell deposit on the section 3 (or 2) on the plate. (He uses
    a cinerator while we are using Bunsen burner to sterilize the loop.)

    * There are many different ways to streak plates to achieve the


    goal of depositing single cells on the plate. We are going
    to use the T-streak plate technique.

    Fall 2020 Microbiology Lab #3


    T-Streak Pattern
    Practice streaking patterns on a
    paper.
    1. Draw a circle and draw a T to divide
    the circle into 3 sections.
    2. Use a pen to draw T-Streak streaking
    pattern inside the circle.
    3. Pretending that the circle is your plate
    and the pen is your inoculating loop,
    “flame” your loop on a “imaging
    burner” between each streak.

    Fall 2020 Microbiology Lab #3


    Technique Review
    1. How many times do you flame your loop when you
    perform the T-streak technique?
    2. What would happen if you forget to flame your loop
    between the streaks?
    3. How many times do you need to sample the culture
    to do the T-streak?
    4. Explain why it is necessary to incubate plates up-
    side-down.

    Fall 2020 Microbiology Lab #3


    Lab Assignment #2
    1. Draw a T on the back of a TSA plate and mark
    the three area with 1,2,3.
    2. Label the edge of your plate with Sec#, Seat#, Initials, Date, Culture
    name
    3. Holding your loop, practice the T-streak pattern on the back of a
    plate.
    (Make sure that your streak closely without overlapping.)
    Glide the loop across the plate gently .
    Flame your loop between streaks.
    4. When your are ready, streak a mixed bacterial suspension assigned
    to you on the Plate using the T-streak technique. (See the streaking
    technique on the demo video.)
    5. Incubate your plate UP-SIDE-DOWN at 37°C for 24 h.

    Fall 2020 Microbiology Lab #3


    What should you expect on your
    streaked plate?

    • The bacterial mixture contains


    one G(+) and one G(-) bacteria.
    • You should see growth of two
    different bacteria (different
    colonial morphology) on your
    plate after incubation.
    • You should see isolated
    bacterial colonies in the section
    2 or 3.

    Fall 2020 Microbiology Lab #3

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