Revision as of 12:26, 25 August 2008

Construction of pFlgA - GFP Generator

Aim : Construction of "pFlgA-RBS-GFP-dbl ter" (pFlgA-E0430/E0432)

Ligation name	Description	Antibio	Number Colonies observed	Fluorescence	Comments
Ligations
L158	D110(BV) - D131(BI) rbs-TetR - gfp tripart	Amp	-	-	to do again
L159	D125(FV) - D109(FI) rbs-LasI - double terminator	Kan	-	-	to do again
L162	D107(BV) - D163(BI) rbs-LasI - gfp generator	Amp	-	-	to do again
L163	D110(BV) - D163(BI) rbs-TetR - gfp generator	Amp	-	-	to do again
Controls
TL158	D110(BV)	Amp	-	-	to do again
TL159	D125(FV)	Kan	-	-	to do again
TL162	D107(BV)	Amp	-	-	to do again
TL163	D110(BV)	Amp	-	-	to do again
Positive Control	pUC19	Amp	358 (efficiency 3.5.10^7)	No	OK

=> We decided to do again the digestions of the ligation's reactions

@@ Line 108: / Line 108: @@
 =='''Digestion'''==
 [[Team:Paris/Notebook/Protocols#Digestion |Protocol]]
+{| border="1" style="text-align: center"
+|'''Name'''
+|'''Template DNA'''
+|'''Description'''
+|'''Vol MP (µl)'''
+|'''Vol H2O (µl)'''
+|'''Enzymes'''
+|-
+|D107
+|MP105.1
+|rbs-LasI (BV)
+|14.7 (0.5µg of plasmid)
+|11
+|SpeI / PstI
+|-
+|D109
+|MP105.1
+|rbs-LasI (FI)
+|14.7
+|11
+|EcoRI / SpeI
+|-
+|D110
+|MP106
+|rbs-TetR (BV)
+|9
+|15.7
+|SpeI / PstI
+|-
+|D125
+|MP118.1
+|Double terminator (FV)
+|4.54
+|20.16
+|EcoRI and XbaI
+|-
+|D163
+|MP143
+|gfp generator  (BI)
+|6.66
+|18.04
+|XbaI and PstI
+|}