Tokyo/Preliminary assays
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Revision as of 12:11, 20 October 2007
Preliminary assays
Contents |
AHL preparation
OD correction
Date:
2007/09/21
Purpose:
Samples:
Procedure:
Result:
Conclusion:
Wash
Date:
20070925
Purpose:
Samples:
Procedure:
Result:
Conclusion:
Detailed AHL assay
Date:
20070925
Purpose:
Samples:
Procedure:
Result:
Conclusion:
New-AHL assay
Date:
20070926
Purpose:
Samples:
Procedure:
Result:
Conclusion:
Lux-lac hybrid promoter Check
Date:
2007/09/20, 21
Purpose 1:
To check if LacI hybrid promoter is activated by AHL and repressed by LacI.
Purpose 2:
To obtain parameters of LacI hybrid promoter for computational simulation.
Samples:
1. pTrc99A + [LacI hybrid promoter – GFP] (LacI+)
2. pBR322 TetR + [LacI hybrid promoter – GFP] (lacI-)
3. pTrc99A + [placQI – GFP] (placQI is constitutive promoter)
4. pTrc322 TetR + [LacI promoter – GFP]
5. pBR322 TetR + [LacI promoter – GFP]
Repressor LacI expression:
pTrc99A expresses LacI
pBR322 TetR does NOT express LacI
Antibiotics resistance:
pTrc99A gives ampicillin-resistance
pBR322 TetR gives ampicillin-resistance
[LacI hybrid promoter – GFP] gives kanamicin-resistance
[placQI – GFP] gives kanamicin-resistance
