IGEM:IMPERIAL/2006/Protocols/S01656new
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New Protocal to test aiiA Production
Outline
- 9am - Make fresh day culture
- 11am - Add known AHL concs + IPTG
- Wait 7 hrs
- 6pm
- Test Flourescence
- Spin down cells + collect supernatant
- The Next Day - Test frozen supernatant for AHL with T9002
- This will show you that aiiA is being produced and that the cells flouresce.
- This can be easily adapted to mesure AHL conc every hour so that you get a curve. however we will not need to do this for this aiiA part as the aiiA gene is untagged and may have diferent properties to the aiiA we will actually be using This test is solely to see if this gene works.
Detailed Protocal
Equipment/Materials
- Falcon tubes or similar
- Photospectrometer
- Gilson pipettes
- Eppendorf tubes
- Centrifuge
- Plain LB medium
- LB medium with 50 μg/mL Kanamycin
- LB medium with 50 μg/mL Ampicillin
- E. coli DH5a strain with part S01656
- E. coli DH5a strain with part T9002
- Perkin Elmer Victor 3 Fluorimeter
- 96 well plate
