The Tube-to-Tube Transfer Technique

The text of the narration (read by the Apple Macintosh OS 9 "Victoria, high-quality" computer voice) is as follows:

We will be transferring organisms from the culture in the green-plugged tube to the sterile broth medium in the red-capped tube.

The inoculating loop is held in the flame until the entire wire is red-hot. This sterilizes the wire and part of the handle. Note that the instrument is held at an angle and not horizontally.

It is always a good idea to loosen the caps and plugs before the transfer operation begins. The tubes are held at an angle and the caps and plugs are pulled out (as shown) with the hand that is holding the inoculating loop.

Briefly flame the open ends of the tubes. Dip the sterile loop into the culture and transfer the inoculum to the broth tube, again by simply dipping the loop into the liquid.

The loop still contains organisms. It is always a good idea to run the loop against the glass on the way out of the inoculated tube in order to break the film that has formed across the loop. This lessens the probability of an aerosol if the film across the loop should break when outside the tube.

Briefly flame the tubes again and apply the plugs and caps which have been held in the same hand throughout the procedure.

Carefully flame the loop, starting at the base of the wire. By the time the flame reaches the tip, the organisms have dried out and died off, and no aerosol of living cells is possible.

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Page content was last modified on 6/18/14 at 2:00 PM, CDT.
John Lindquist, Department of Bacteriology,
University of Wisconsin – Madison