Streptococcus Bacteriocin (Dual Layer) Assays
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Goal
- Set up three different assays to test a bacteriocin's killing ability.
Protocol
Day 1: Plating S. pneumoniae
1. Grab a frozen cozy from the freezer in the MEE Lab and go to the -80°C freezer next to the MEE Lab.
2. Grab the containers of producer and target strains from the freezer and place them in the cozy
3. While under the hood:
- a. Have a beaker of Virkon ready for disposing inoculating loops
- b. Label two TSB agar blood plates “<producer name>” and “<target name>”
- c. Open tube of producer strain and grab a chunk of ice containing the strain using an inoculating loop
- d. Grab the “<producer name>” plate and streak the loop horizontally, from top to bottom, across the top-third of the agar
- e. Put the inoculating loop in Virkon
- f. Grab a new inoculating loop and start streaking from the right side of the last streak and proceed to streak the bacteria downwards across the right half of the plate
- g. Put the inoculating loop in Virkon
- h. Grab a new inoculating loop and start streaking from the bottom side of the last streak and proceed to streak the bacteria upwards across the left half of the plate
- i. Put the inoculating loop in Virkon
Repeat Steps c-i for the target strain on the “<target name>” plate Put plates in 37°C 5% CO2 incubator overnight.
Day 2: Preparation
Growing Producer and Target Strains