Performed chlorine dioxide gas exposure to the Bacillus thuringiensis spores on the glass cover slips in the 50 mL conical tubes as performed previously (1-31-14).
Three different chlorine dioxide concentrations were weighed out using the analytical balance in the Pruess Lab for a combined mass of: 25 mg, 50 mg, and 100 mg. The two components to generate the gas, Part A and Part B, are combined in a 1:1 ratio in the 50 mL tube to generate the gas. So, in order to get the combined mass, a 12.5 mg, 25 mg, and 50 mg masses were weighed out for each of the two components.
Parts A and B were combined in the 50 mL conical tubes, the two mix at the bottom. Quickly after the two are combined, the glass cover slips containing the dried spores were aseptically added to the 50 mL tube. Due to the size of the cover slip and the narrowing of the tube the glass never comes in direct contact with the two components at the bottom. After the addition of the cover slip, the tube is closed and sealed with Parafilm.
The tubes were transferred to the small shaking incubator protected from light. The tubes were gently rocked using the shaker, but no additional heat was applied. The tubes were incubated at room temperature, gently rocking, and protected from light for six hours.
After six hours, the glass cover slips were removed from the 50 mL conical tubes inside a fume hood and transferred to a new 50 mL tube containing 1.0 mL of sterile ddH2O. During this aseptic transfer, the glass cover slips were all broken using the forceps. This process will allow the glass containing the dried spores to come in better contact with the water and promote re-suspension. The tubes containing the broken glass were vortexed for 10 seconds and allowed to incubate at room temperature for 5 minutes.
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| Broken glass cover slips in 1 mL H2O inside a 50 mL tube |
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