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-	== [[Tokyo/AHL preparation |AHL preparation]] ==	+	__NOTOC__
-	== AHL assay==	+	<br>[[Tokyo/Works|Works top]]&nbsp;&nbsp;&nbsp;0.[[Tokyo/Works/Hybrid promoter|Hybrid promoter]]&nbsp;&nbsp;&nbsp;1.[[Tokyo/Works/Formulation |Formulation]]　　'''2.[[Tokyo/Works/Assay |Assay1]]'''&nbsp;&nbsp;&nbsp;3.[[Tokyo/Works/Simulation |Simulation]]&nbsp;&nbsp;&nbsp;4.[[Tokyo/Works/Assay2 |Assay2]]&nbsp;&nbsp;&nbsp;5.[[Tokyo/Works/Future works |Future works]]
-		+	<br><br>
-	===Date: ===	+	[[Tokyo/Hill function fitting |Objective of this assay]]&nbsp;&nbsp;&nbsp;[[Tokyo/AHL assay|Effect of AHL]]&nbsp;&nbsp;&nbsp;[[Tokyo/IPTG assay|Effect of LacI-IPTG]]　　[[Tokyo/Preliminary assays |Preliminary assays]]
-	2007/09/06, 07	+
-	===Purpose1: ===	+
-	To practive AHL assay.	+
-	===Purpose2: ===	+
-	To check if AHL works.	+
-	===Samples: ===	+	== ==
-	<br>luxR + GFP in DH5a (AHL-activated promoter)	+	'''Preliminary assays were conducted'''
-	<br>placQI + GFP in DH5a (pos. con.)	+	<br>- '''to practice and experience the assays with AHL and IPTG'''
-	<br>pSBΔP+ GFP (neg. con.)	+	<br>- '''to check if and how our materials actually worked.'''
-	<br>ΔP = promoter deleted	+	== [[Tokyo/AHL preparation |AHL preparation]] ==
-		+	== [[Tokyo/Experimental precision and accurecy|Experimental precision and accuracy]] ==
-	===Procedure: ===	+	== [[Tokyo/AHL assay practice|AHL assay practice]] ==
-	<br>The final concentration of AHL is 6 pM.	+	== [[Tokyo/Lux-lac hybrid promoter activation check|Lux-lac hybrid promoter activation check]] ==
-		+	== [[Tokyo/OD correction|OD correction]] ==
-	===Result: ===	+	== [[Tokyo/Wash|Wash]] ==
-	===Conclusion: ===	+	== [[Tokyo/New AHL check|New AHL check]] ==
-	6pM of AHL is not enough to activate the AHL-activated luxR promoter.	+	<!--== [[Tokyo/Hybrid promoter activation & repression check|Hybrid promoter activation & repression check ]] ==
-		+
-	== AHL assay==	+
-		+
-	===Date: ===	+
-	2007/09/15	+
-	===Purpose: ===	+
-	check if lux-lac hybrid promoter works	+
-		+
-	===Samples: ===	+
-	<br>Lux-lac hybrid promoter (before blue light check)	+
-	<br>Lux-lac hybrid promoter (after blue light check)	+
-	<br>placQI (pos. con.)	+
-	<br>modified A4 (promoter deleted) (neg. con.)	+
-		+
-	===Procedure: ===	+
-	[[Tokyo/AHL assay Standard protocol |AHL assay Standard protocol ]]	+
-		+
-	===Result: ===	+
-		+
-	===Conclusion: ===	+
-		+
-	== OD correction ==	+
-		+
-	===Date: ===	+
-	2007/09/21	+
-	===Purpose: ===	+
-	===Samples: ===	+
-	<br>ΔP-GFP in pTrc99A (neg. con.)	+
-	<br>placQI in pTrc99A (pos. con.)	+
-	<br>Only LB (blank)	+
-		+
-	===Procedure: ===	+
-	===Result: ===	+
-	===Conclusion: ===	+
-		+
-		+
-	== Wash ==	+
-		+
-	===Date: ===	+
-	20070925	+
-	===Purpose: ===	+
-	To determine the actual fluorescence of GFP expressed under the promoters by removing other influencial facters such as culture media.	+
-		+
-	===Samples: ===	+
-	<br>A4 placQI in pTrc99A (pos. con.)	+
-	<br>A4ΔP in pTrc99A	+
-	<br>A4 hybrid promoter in pBR322	+
-		+
-	===Procedure: ===	+
-	[[Tokyo/AHL assay Standard protocol |AHL assay Standard protocol ]]	+
-	<br>[[Tokyo/Wash |Wash ]]	+
-		+
-	===Result: ===	+
-	===Conclusion: ===	+
-		+
-		+
-	== Detailed AHL assay ==	+
-		+
-	===Date: ===	+
-	20070925	+
-	===Purpose: ===	+
-	<br>To determine the AHL dependent behaviour in narrower range of AHL concentration for the analyses and simulation.	+
-		+
-	===Samples: ===	+
-	===Procedure: ===	+
-	===Result: ===	+
-	===Conclusion: ===	+
-		+
-	== New-AHL assay ==	+
-		+
-	===Date: ===	+
-	20070926	+
-	===Purpose: ===	+
-	<br>To determine and compare the activity of the AHL used so far and that of the newly perchased AHL.	+
-		+
-	===Samples: ===	+
-	<br>A4 placQI in pTrc99A (pos. con.)	+
-	<br>A4ΔP in pTrc99A	+
-	<br>A4 hybrid promoter in pBR322	+
-		+
-	===Procedure: ===	+
-	<br>The old and the newly delivered AHL was used.	+
-	<br>[[Tokyo/AHL assay Standard protocol |AHL assay Standard protocol ]]	+
-	<br>The final concantrations of AHL applied here were 0.01, 0.1, 0.5, 1, 10, 100, and 1000 nM.	+
-		+
-	===Result: ===	+
-	===Conclusion: ===	+
-		+
-	== Lux-lac hybrid promoter Check ==	+
-		+
-	[[Image:luxlachybrid.jpg]]	+
-		+
-	=== 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: ===	+
-	<br>1. pTrc99A + [LacI hybrid promoter – GFP] (LacI+)	+
-	<br>2. pBR322 TetR + [LacI hybrid promoter – GFP] (lacI-)	+
-	<br>3. pTrc99A + [placQI – GFP] (placQI is constitutive promoter)	+
-	<br>4. pTrc322 TetR + [LacI promoter – GFP]	+
-	<br>5. pBR322 TetR + [LacI promoter – GFP]	+
-		+
-	<br>'''Repressor LacI expression:'''	+
-	<br>pTrc99A expresses LacI	+
-	<br>pBR322 TetR does NOT express LacI	+
-		+
-	<br>'''Antibiotics resistance:'''	+
-	<br>pTrc99A gives ampicillin-resistance	+
-	<br>pBR322 TetR gives ampicillin-resistance	+
-	<br>[LacI hybrid promoter – GFP] gives kanamicin-resistance	+
-	<br>[placQI – GFP] gives kanamicin-resistance	+
-	===Procedure:===	+	<br>
-	<br>[[Tokyo/AHL assay Standard protocol |AHL assay Standard protocol ]]	+	-->

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Latest revision as of 05:12, 27 October 2007

Works top   0.Hybrid promoter   1.Formulation　　2.Assay1   3.Simulation   4.Assay2   5.Future works

Objective of this assay   Effect of AHL   Effect of LacI-IPTG　　Preliminary assays

Preliminary assays were conducted
- to practice and experience the assays with AHL and IPTG
- to check if and how our materials actually worked.

AHL preparation

Experimental precision and accuracy

AHL assay practice

Lux-lac hybrid promoter activation check

OD correction

Wash

New AHL check