Name:

Section:

Aseptic Technique
(tube to tube transfer of inoculum)

Objective: Transfer of microorganisms from test tube to test tube using aseptic technique while
preventing contamination along the transfer process.

1.Holding the 2 test tubes on your hands, sterilize the loop.

Sterilization of inoculating loop: Tilt at 45 degrees. Sterilize till the wire is heated (red hot color). Cool
down for 1 minute.

2. Open the test tube labeled as test tube #1 and heat the rim, while holding the cotton plug. Maintain
sterility of the wire in inoculating loop before acquiring the inoculum.

Manipulation technique: Inner part of the cotton plug does not touch the table, or your fingers. Spaces
in between fingers is meant to hold firmly the outer part of the cotton plug. Cotton plug should be held
correctly.

3. Insert the & gently swirl the loop to get the inoculum. Only the sterile part of the loop can enter the
test tube.

Inoculum: organism found in the medium

Inocula: plural form of inoculum
Medium used in set-up: non-sterile water from faucet

4. Heat the tube rim and seal the test tube #1 with the cotton plug.

5. Open the test tube labeled as test tube #2 and heat the rim, while holding the cotton plug.

6. Insert & gently swirl the loop to transfer inoculum to tube #2.

7. Heat the tube rim and seal the test tube #2 with the cotton plug.

8. Decontamination: heat the inoculating loop. Cool down for 1 minute.

Procedure Prepared by:

Diana Verano