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Lab Meeting 09/20/2013

Lab Meeting 09/20/2013. Catherine Doyle. Caffeine Derivative Experiment Results. eCDM8. c.Dog RBS. GFP. psB1A8. High Copy. TetA. psB4C5. Low Copy. eCDM8. c.Dog RBS. GFP. psB1A8. High Copy. AdhE. psb4C5. High Copy. No AdhE - plasmid. No TetA gene. AdhE - cells.

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Lab Meeting 09/20/2013

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  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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