17 June 2008
- Our goal for the day was to make different concentrations of our bacterial cultures of E. coli strain SM10, plasmid PVJ4 containing SRRz fragment from RY100 cloned into pBAD18mob.1. We then added FastLyse in order to lyse the cells and then record resistance measurements. We centrifuged six test tubes various times with 10 mL each of culture and then resuspended with distilled water.
- Tube 1: Centrifuged once and resuspended in 10 mL Distilled water
- Tube 2: Centrifuged twice and resuspended in 10 mL Distilled water
- Tube 3: Centrifuged 3X, resuspended in 10 mL Distilled water
- Tube 4: Centrifuged 4x, resuspended
- Tube 5: Centrifuged 5x, resuspended
- Tube 6: Centrifuged 6x, resuspended
- Using a 12-well plate, we added 3mL of bacteria and 330 microliters of FastLyse to 6 wells and distilled water to a seventh well.
- After adding the FastLyse to the distilled water, the change in resistance was greater than 700 kilo Ohms.
- The resistance measurements of the 6 bacterial solutions before the addition of FastLyse were all within 260 -369 kilo Ohms. After addition of the FastLyse, resistance drops were all very similar and could only be attributed to the addition of FastLyse and corresponding electrolytes and ions instead of lysis.
- Our next goal is to perform lysis on different concentrations of bacteria using the Freeze-Thaw method.
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