Kirby-Bauer Technique

Today in lab we used the Kirby-Bauer Technique to make a streak plate.

First of all we used Mueller-Hinton Agar plates, which are used to test the antibiotics (abx) susceptibility to certain bacteria.
here are the steps used before and during a KB process:

1. Put 1mL of sterile broth into a sterile test tube using your best aseptic technique
2. Then, find your streak plate from the hand swiping lab we've done in the past.
3. Using a bunsen burner and an IN. loop heat the loop until it is red hot and then allow it to cool for a few seconds
4. Use the heat cleaned loop to take 1 isolated colony from the streak plate, and stir it into the sterile broth in your test tube.
5. Once you have accomplished this you need to use the Kirby-Bauer Technique:

*Get your self a MH agar plate (actually 2, because you're making 2)
*Find a sterile swab
*Work close to the fire in order to help prevent contamination
*Dip the sterile swab into the bacteria broth in the test tube and make sure it is very wet
*Streak the swab across the MHA plate back and forth making sure to overlap your streaks. Make a "lawn"
- There should be no open gaps.
*Go about 3/4 of the way down, turn the plate 90 degrees, and continue (repeat 2X)
*After you have a "lawn" of bacteria on the plate close the lid
*Find yourself 5 abx disks and a no control disk
*Using flame sterilized tweezers put 3 disks onto each plate spread out enough that they will not overlap at all.
*Label them on the bottom of the plate.
*Put your plate into the insulator and wait for some pretty colors!! :)

Have Fun!

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