Chemical Hygiene Plan and Safety Manual - LP3013.4 Aseptic Technique

8.3.5.4 Open a sterile box of pipette tips and slowly pipette the appropriate volume into the labeled test tube, replacing the cap immediately after transfer.

8.3.5.4.1 Never place the pipette with the tip attached on the counter.

8.3.5.5 Repeat for remaining dilutions using a new pipette tip with every solution.

8.3.6 To streak samples onto a slant tube:

8.3.6.1 Label a sterilized test tube with sample identification and test type.

8.3.6.2 After preparing the work space as outlined in section 8.2, sanitize hands and don gloves.

8.3.6.3 Open the sample bag.

8.3.6.4 Open a sterile plastic loop, and insert the loop into the sample bag.

8 3 6 5 Open the cap of the test tube, and streak the loop on the surface of the slanted agar Ensuring to do so quickly and evenly in a zig-zag pattern

8 3 6 5 1 Do not allow the loop to puncture the agar surface

8 3 6 5 2 Never place the loop on the counter

8 3 6 6 After streaking, replace the cap quickly and dispose of gloves in trash and dispose of loop in autoclave bags

8 3 6 7 Repeat with remaining samples ensuring to use a new loop for each sample and tube

8 3 7 To inoculate the butt or stab of test tube:

8 3 7 1 Label a sterilized test tube with sample identification and test type

8 3 7 2 After preparing the work space as outlined in section 8 2, sanitize hands and don gloves

8.3.7.3 Open the petri-dish containing colonies to be used for inoculation.

8.3.7.4 Open a sterile plastic loop, and collect 1 colonies (1 loopful.)

8.3.7.5 Open the cap of the test tube, and stab the loop into the middle of the surface of the agar, refer to the corresponding test method to determine the depth the loop needs to go into the agar. Stabs will go down part-way and butt inoculations will go further towards the bottom of the tube.

8.3.7.5.1 Do not allow the loop to touch the sides of the test tubes.

8.3.7.5.2 Never place the Loop on the counter.

8.3.7.6 After inoculation, replace the cap quickly and dispose of gloves in trash and dispose of loop in autoclave bags.

8 3 7 7 Repeat with remaining samples ensuring to use a new needle for each sample and tube

8 3 8 To transfer samples to petri dishes:

8 3 8 1 Follow the diagram in figure one to follow the proper T-streak pattern

8 3 8 2 Label a sterilized petri dish with sample identification and test type