From 2007.igem.org

Preliminary assays 

Contents 1 AHL preparation 2 OD correction 2.1 Date: 2.2 Purpose: 2.3 Samples: 2.4 Procedure: 2.5 Result: 2.6 Conclusion: 3 Wash 3.1 Date: 3.2 Purpose: 3.3 Samples: 3.4 Procedure: 3.5 Result: 3.6 Conclusion: 4 Detailed AHL assay 4.1 Date: 4.2 Purpose: 4.3 Samples: 4.4 Procedure: 4.5 Result: 4.6 Conclusion: 5 New-AHL assay 5.1 Date: 5.2 Purpose: 5.3 Samples: 5.4 Procedure: 5.5 Result: 5.6 Conclusion: 6 Lux-lac hybrid promoter Check 6.1 Date: 6.2 Purpose 1: 6.3 Purpose 2: 6.4 Samples: 6.5 Procedure:

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

Procedure: