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Bacterial cell culture: Difference between revisions
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→Procedure: Removed suggestion to eject pipette tip into culture. This is poor sterile technique.
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John Wertz (talk | contribs) (→Procedure: Removed suggestion to eject pipette tip into culture. This is poor sterile technique.) |
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#Flame a glass pipette, open the bottle of medium and flame the mouth, measure out the amount you need to fill your tubes, flame the cap and recap the bottle as quickly as possible. | #Flame a glass pipette, open the bottle of medium and flame the mouth, measure out the amount you need to fill your tubes, flame the cap and recap the bottle as quickly as possible. | ||
#Remove the tube cap, flame the top of the culture tube, pipette in 5 ml, flame the top of the tube, and cap it. | #Remove the tube cap, flame the top of the culture tube, pipette in 5 ml, flame the top of the tube, and cap it. | ||
#Pick up one colony by tapping a small (0.1 μl) pipette tip (held on a pipette) on the surface of the plate. Uncap a tube, flame the top, | #Pick up one colony by tapping a small (0.1 μl) pipette tip (held on a pipette) on the surface of the plate. Uncap a tube, flame the top, tip the tube so as to transfer cells from the pipette tip to the surface of the media without touching the inside of the tube with the non-sterile portion of the pipetter, flame, cap. You can also use a sterile toothpick for transfer. | ||
#*Often people use just a sterile metal loop (sterile by flaming) to place the colony in the tube. This is because flaming assures the sterility of the loop, whereas disposables such as pipette tips and toothpicks can be contaminated, and cannot be flamed on the spot. | #*Often people use just a sterile metal loop (sterile by flaming) to place the colony in the tube. This is because flaming assures the sterility of the loop, whereas disposables such as pipette tips and toothpicks can be contaminated, and cannot be flamed on the spot. | ||
#Pipette the desired amount of [[Endy:Preparing Antibiotic Stocks|antibiotic]] into each tube along the wall. Do not put the non-sterile part of the pipette inside the tube and use a new tip for each tube. | #Pipette the desired amount of [[Endy:Preparing Antibiotic Stocks|antibiotic]] into each tube along the wall. Do not put the non-sterile part of the pipette inside the tube and use a new tip for each tube. | ||
