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Caffeine Derivative Experiment Results Analysis: eCDM8.c Dog RBS GFP psB1A8 AdhE psB4C5 TetA

Dive into the findings of a caffeine derivative experiment showcasing results from eCDM8.c, Dog RBS, and GFP psB1A8, together with TetA resistance factors. Explore the impact of AdhE psB4C5, high-copy plasmids, and the absence of AdhE in cells, with TetA additions and growth conditions. Delve into methods for testing TetA resistance, considerations for controls, liquid medium versus plates, and optimal growth temperatures. Join the discussion on experimental setups and potential variables for enhanced outcomes.

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Caffeine Derivative Experiment Results Analysis: eCDM8.c Dog RBS GFP psB1A8 AdhE psB4C5 TetA

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Presentation Transcript

  1. Lab Meeting 09/20/2013 Catherine Doyle

  2. Caffeine Derivative Experiment Results

  3. eCDM8 c.Dog RBS GFP psB1A8 High Copy

  4. TetA psB4C5 Low Copy

  5. eCDM8 c.Dog RBS GFP psB1A8 High Copy

  6. AdhE psb4C5 High Copy

  7. No AdhE- plasmid No TetA gene AdhE- cells AdhE- cells TetA added No TetA added TetA added No TetA added

  8. No AdhE- plasmid No TetA gene AdhE- cells AdhE- cells TetA added No TetA added TetA added No TetA added Grow at 37C O/N Repeat for 36 Days: Remove 100ul from each tube 200ul for Cell Density and GFP Expression 3X 100ul in 4.9ml of new LB 100ul on plate

  9. No AdhE- plasmid No TetA gene AdhE- cells AdhE- cells TetA added No TetA added TetA added No TetA added Grow at 37C O/N Repeat for 36 Days: Remove 100ul from each tube 200ul for Cell Density and GFP Expression 3X 100ul in 4.9ml of new LB Increase TetA 0.005ul every three days For plasmids with TetA resistance (Will have to experimentally determine. 100ul on plate

  10. Ruling Out False Positives from TetA Plate on LB Amp/Chlor Plate on LB Amp/Chlor/TetA

  11. Questions? • Whether we need a control where cells lacking tetA gene are treated the same way? • Whether we need a control where adhE-cells are lacking the adhE plasmid? • Is liquid better than plates? • Should we let them grow at 37C or room temperature?

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