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Line 16:		Line 16:
	| style="background: #ccffcc;" |MP15		| style="background: #ccffcc;" |MP15
	|Att B		|Att B
-	|Sigma Att B rev within pSB2K3	+	|Lambda Att B rev within pSB2K3
	|		|
	| [[Paris/August 30| August 30]]		| [[Paris/August 30| August 30]]
Line 22:		Line 22:
	| style="background: #ccffcc;" |MP16		| style="background: #ccffcc;" |MP16
	|Att P		|Att P
-	|Sigma Att P within pSB2K3	+	|Lambda Att P within pSB2K3
	|		|
	| [[Paris/August 30| August 30]]		| [[Paris/August 30| August 30]]
Line 42:		Line 42:
	|Lambda Att B rev within pSB2K3		|Lambda Att B rev within pSB2K3
	|		|
-	|[[Paris/August 22| August 22]]	+	|[[Paris/August 30| August 30]]
	|- style="background: #cccccc;"		|- style="background: #cccccc;"
	| style="background: #ccffcc;" |S47		| style="background: #ccffcc;" |S47
Line 48:		Line 48:
	|Lambda Att P within pSB2K3		|Lambda Att P within pSB2K3
	|		|
-	|[[Paris/August 22| August 22]]	+	|[[Paris/August 30| August 30]]
		+	|}
	== Digestions ==		== Digestions ==
		+	we digest:
-	== Growth kinetic on w121 strains ==	+	* MP1.1 as Bw vector with Spe1 & Pst1
-		+	* MP15 as Bw insert with Xba1 & Pst1
-	In order to precise the growth behavior/kinetics of our strain W121 in smaller DAP concentration, we fit those last ones in function of our last kinetics results (06/08/07)	+	* MP16 as Bw insert with Xba1 & Pst1
-		+
-	{{Paris_KineticArray| Title = w121 kinetic as a function of DAP and supplemented medium (S0.x)	+
-	|A1=H<sub>2</sub>0|A2=H<sub>2</sub>0|A3=H<sub>2</sub>0|A4=H<sub>2</sub>0	+
-	|A5=H<sub>2</sub>0|A6=H<sub>2</sub>0|A7=H<sub>2</sub>0|A8=H<sub>2</sub>0	+
-	|A9=H<sub>2</sub>0|A10=H<sub>2</sub>0|A11=H<sub>2</sub>0|A12=H<sub>2</sub>0	+
-	|B1=H<sub>2</sub>0	+
-	|B2=LB+0µM DAP|B3=LB+5µM DAP|B4=LB+8µM DAP|B5=LB+10µM DAP|B6=LB+12µM DAP	+
-	|B7=LB+15µM DAP|B8=LB+17µM DAP|B9=LB+20µM DAP|B10=LB+30µM DAP|B11=LB+40µM DAP	+
-	|B12=H<sub>2</sub>0|C1=H<sub>2</sub>0	+
-	|C2=S0.2+0µM DAP|C3=S0.2+5µM DAP|C4=S0.2+6µM DAP|C5=S0.2+7µM DAP|C6=S0.2+8µM DAP	+
-	|C7=S0.2+9µM DAP|C8=S0.2+10µM DAP|C9=S0.2+15µM DAP|C10=S0.2+17µM DAP|C11=S0.2+20µM DAP	+
-	|C12=H<sub>2</sub>0|D1=H<sub>2</sub>0	+
-	|D2=S0.4+0µM DAP|D3=S0.4+2µM DAP|D4=S0.4+3µM DAP|D5=S0.4+4µM DAP|D6=S0.4+5µM DAP	+
-	|D7=S0.4+8µM DAP|D8=S0.4+10µM DAP|D9=S0.4+12µM DAP|D10=S0.4+15µM DAP|D11=S0.4+20µM DAP	+
-	|D12=H<sub>2</sub>0|E1=H<sub>2</sub>0	+
-	|E2=S0.6+0µM DAP|E3=S0.6+1µM DAP|E4=S0.6+2µM DAP|E5=S0.6+3µM DAP|E6=S0.6+4µM DAP	+
-	|E7=S0.6+5µM DAP|E8=S0.6+6µM DAP|E9=S0.6+7µM DAP|E10=S0.6+8µM DAP|E11=S0.6+10µM DAP	+
-	|E12=H<sub>2</sub>0|F1=H<sub>2</sub>0	+
-	|F2=S0.8+0µM DAP|F3=S0.8+0,5µM DAP|F4=S0.8+1µM DAP|F5=S0.8+2µM DAP|F6=S0.8+3µM DAP	+
-	|F7=S0.8+4µM DAP|F8=S0.8+5µM DAP|F9=S0.8+6µM DAP|F10=S0.8+7µM DAP|F11=S0.8+10µM DAP	+
-	|F12=H<sub>2</sub>0|G1=H<sub>2</sub>0	+
-	|G2=H<sub>2</sub>0|G3=H<sub>2</sub>0|G4=H<sub>2</sub>0|G5=H<sub>2</sub>0|G6=H<sub>2</sub>0	+
-	|G7=H<sub>2</sub>0|G8=H<sub>2</sub>0|G9=H<sub>2</sub>0|G10=H<sub>2</sub>0|G11=H<sub>2</sub>0	+
-	|G12=H<sub>2</sub>0|H1=H<sub>2</sub>0|H2=H<sub>2</sub>0|H3=H<sub>2</sub>0|H4=H<sub>2</sub>0	+
-	|H5=H<sub>2</sub>0|H6=H<sub>2</sub>0|H7=H<sub>2</sub>0|H8=H<sub>2</sub>0	+
-	|H9=H<sub>2</sub>0|H10=H<sub>2</sub>0|H11=H<sub>2</sub>0|H12=H<sub>2</sub>0}}	+
-	The 96 well plate is surrounded by H<sub>2</sub>O in order to maintain humidity during the assay. In each slot, 200µL of the growth medium (LB or S0.x) is mixed with 2µL of w121 culture grown ON, and with different amount of DAP (see table).	+	Protocol:<br>
-	The growth profile is measured over a period of 20H, a DO measurement is acquired each 4min10s.	+	ADN: 20µL<br>
		+	BSA: 0.5µL<br>
		+	NE Buffer 2: 5µL<br>
		+	Enzyme 1: 2µL<br>
		+	Enzyme 2: 2µL<br>
		+	H2O: 20.5µL<br>

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Latest revision as of 12:02, 31 August 2007

yesterday -- tomorrow

Contents 1 Expression test of DapA promotor by differential DAP repression 2 Plasmids: MiniPrep Products 3 Strains (glycerol stock) 4 Digestions

Expression test of DapA promotor by differential DAP repression

Plasmids: MiniPrep Products

Plasmids: MiniPrep Products
Number	Name	Description	Products in use	Date
MP15	Att B	Lambda Att B rev within pSB2K3		August 30
MP16	Att P	Lambda Att P within pSB2K3		August 30

Strains (glycerol stock)

Strains glycerol stock
Number	Name	Description	Verified clone	Date
S46	MP15	Lambda Att B rev within pSB2K3		August 30
S47	MP16	Lambda Att P within pSB2K3		August 30

Digestions

we digest:

• MP1.1 as Bw vector with Spe1 & Pst1
• MP15 as Bw insert with Xba1 & Pst1
• MP16 as Bw insert with Xba1 & Pst1

Protocol:
ADN: 20µL
BSA: 0.5µL
NE Buffer 2: 5µL
Enzyme 1: 2µL
Enzyme 2: 2µL
H2O: 20.5µL