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BISC209: Aseptic Transfer: Difference between revisions
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→Aseptic Transfer
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1. Label the destination container for the culture. | 1. Label the destination container for the culture. | ||
2. Holding your loop like a pencil, insert the loop into the flame as illustrated in Figure | 2. Holding your loop like a pencil, insert the loop into the flame as illustrated in Figure 1. The orientation of the loop wire in the flame is important for proper incineration. Keep the wire in the flame until it is red-hot. Then move the adjacent nonwire part of the loop lightly through the flame. The wire will now be sterile, and the nonwire part will have any dust burned off that might have fallen into the media during the transfer procedure. Remember that the loop is now hot and sterile. Allow the loop to cool for a few seconds in the air. <br> | ||
[[Asep1.jpg]]<br> | |||
Figure 1: Proper flaming of a loop. Note how the handle is held by only the thumb and first two fingers and the loop is inserted into the hottest part of the flame. | |||
3. Pick up the stock broth tube of your organism with your other hand, while still holding the sterile loop. With the hand holding the loop, use your little finger against your palm to remove the cover from the culture tube as shown in Figure | 3. Pick up the stock broth tube of your organism with your other hand, while still holding the sterile loop. With the hand holding the loop, use your little finger against your palm to remove the cover from the culture tube as shown in Figure 2. Do not put the cover down on your bench. If the tube was closed with a cotton or plastic plug, lightly pass the lip of the tube through the Bunsen burner to burn off any adhering dust. If the tube was closed with a plastic cap the lip of the tube should be sterile and this flaming is probably unnecessary. Now, insert the loop into the broth, and then remove it, carrying a loopful of culture. Flame the top of the culture tube, replace the tube cover, and return the tube to a rack.<br> | ||
[[Asep2.jpg]]<br> | |||
Figure 2: Transferring a culture. (a) Removal of a tube cap while manipulating a loop; (b) Obtaining inoculum from a broth tube while maintaining sterility of the cap (note cap in hand).<br> | |||
4. Pick up the labeled destination tube or plate. Remove its cover, (flame if appropriate), insert the loop containing the culture into the broth, swirl gently and remove. Replace the cover and set the tube in the rack. Resterilize the loop before putting it down to avoid contamination of the bench with the culture. Be careful here. When the loop has liquid on it, you must insert the loop into the flame slowly to allow the liquid to evaporate rather than boil, which would splatter live bacterial cells all over the bench, your books and you. | 4. Pick up the labeled destination tube or plate. Remove its cover, (flame if appropriate), insert the loop containing the culture into the broth, swirl gently and remove. Replace the cover and set the tube in the rack. Resterilize the loop before putting it down to avoid contamination of the bench with the culture. Be careful here. When the loop has liquid on it, you must insert the loop into the flame slowly to allow the liquid to evaporate rather than boil, which would splatter live bacterial cells all over the bench, your books and you. | ||
5. Repeat steps 2, 3, and 5 for each liquid culture needed. | 5. Repeat steps 2, 3, and 5 for each liquid culture needed. | ||
Slant to Broth, plate colony to broth: | |||
'''Slant to Broth, plate colony to broth:''' | |||
