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    Lab 3A Culture Transfer

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    Lab 3A Culture Transfer

    Uploaded by

    brucehung
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    Copyright:

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

    Lab 3A Culture Transfer

    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 3 Aseptic Transfer,


    Isolation & Simple Stain

    Exp 2 Culture Transfer (using Aseptic


    Technique)
    Exp 3 Isolation of Pure Culture
    Exp 14 Selective and Differential Media
    Exp 7 Smear Prep
    Exp 8 Simple Stain

    Fall 2020 Microbiology Lab #3


    Lab 3A
    Exp 2 Culture Transfer Techniques

    • Aseptic Technique:
    techniques to prevent contamination of your
    cultures as well as surrounding environment
    Two goals:
    (1) Not introducing contamination to your tube/ bottle/
    plate
    (2) Not allow microbes to escape from your
    tube/bottle/plate

    Fall 2020 Microbiology Lab #3


    Aseptic Technique
    Tips:
    • Practice opening and holding the cap
    of a tube (or bottle) with your pinkie
    while holding a loop or needle.
    (Fig 1-4)
    • Hold the tube at a angle to prevent
    entry of mos in air to the tube
    when the tube is open. (Fig. 1-3)
    • Flame the opening of the tube (or
    bottle) briefly on burner after
    removing the cap (Fig 1) and before
    recap the tube (Fig 3).

    Fall 2020 Microbiology Lab #3


    Aseptic Technique
    (1) Not introducing contamination to your
    tube/bottle/plate
    Please note that microbes (mos) are present in air.
    Tips:
    • Flame your loop in the hottest portion
    of burner till red hot Briefly pass the
    lower portion of the handle through
    flame.
    • Never hold two tubes on one hand
    as shown in Fig. 2.1 in lab manual.
    • Keep your tubes on rack when they are not needed.

    • Minimum the time that the plates or tubes are open to air.

    • When streaking a plate, keep the lid on top of the plate.


    Fall 2020 Microbiology Lab #3
    Demo Videos

    #1 Aseptic Technique Tips. (5:56)


    In this video, Dr. Gary Kaiser offer important tips of aseptic
    technique, including how to hold tubes and loops, and how
    to remove the cap from a tube.

    Note: He uses a electrical cinerator instead of Bunsen burner.


    You will use Bunsen burner in our lab. You should burn
    you loop in the hottest portion of a Bunsen burner till it
    turns to red.

    Fall 2020 Microbiology Lab #3


    Demo Videos

    #2
    Aseptic Technique: Inoculating broth tubes, slant tubes, and s
    tab tubes
    . (5:42)

    In this video, Dr. Gary Kaiser demonstrates how to


    (1) transfer a E. coli broth culture to a tube of sterile TSB,
    (2) transfer the E. coli broth culture to an agar slant and streak
    the slant, and
    (3) inoculate the E. coli culture to a agar tube by stabbing.

    Note: He uses a electrical cinerator instead of Bunsen burner


    and you will use Bunsen burner in our lab.
    Fall 2020 Microbiology Lab #3
    Lab 3 Assignment #1
    1. Practice holding an inoculating loop and using your pinkie of the same
    hand to remove the cap of a sharpie. (pretending the sharpie is a push
    cap test tube.) and practice the aseptic culture transfer procedure. (You
    may also practice this technique at home.)

    2. Each student will be give two broth : TSB in a screw capped tube and
    TSB in a push capped tube.
    a. Label both tubes with Lab Section: T1, T2 or W
    (T1 = Tue, 11 AM Lab, T2 = Tue12:30 PM Lab) , Seat # (1-12),
    Initials, an dDate e.g. T1-6-ET-0901 (=Tue Lab 1, seat 6, initials, date)
    b. Aseptically transfer one loopful from the screw cap tube to the push
    cap tube.
    c. Incubate both tubes at 35°C for 24 h and exam both for growth of
    bacteria..
    Both tubes should be free of bacterial growth if you perform the procedure
    correctly,

    Fall 2020 Microbiology Lab #3

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