Team:Paris/July 31

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Revision as of 14:58, 4 August 2008

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Oligonucleotides design

RBS +

RBS -

Creation of 3 oligonucleotides (O140, O141, O142) in order to create two particular plasmids.

Those plasmids are very useful vectors to amplify promoters and to measure their forces using Standard Promoter Units. The principle is easy to understand. Instead of using a big biobrick containing Promoter, RBS, GFP and Terminators, the Biobrick used for the test is only the Promoter. The standard GFP tripart (E0240) is between the biobrick site, it is just between two PstI restriction sites. This plasmid can be used to amplify a promoter because only the promoter is in the biobrick.

Transformation results

Ligation name	Nb of colonies	Description	Volume
L100	0	rbs-TetR <-> ECFP (RBS+LVA+Term)	3+6
L101	(1)	rbs-TetR <-> GFP tripart+strong rbs	3+6
L102	+/- 100	strong RBS (0,6)<-> YFP	5+10
L103	a lot ++	strong RBS (0,6)<-> GFP	5+5
L104	a lot ++	strong RBS (0,6)<-> LasR activator (+LVA)	5+4
L105	a lot ++	strong constitutive promoter <-> ECFP	5+8
L106	a lot ++	strong constitutive promoter <-> GFP tripart	5+8
L107	a lot ++	strongest constitutive promoter <-> ECFP	5+8
L108	+/- 100 (2 dishes!?)	strongest constitutive promoter <-> GFP tripart	5+8
L109	+/- 30 or a lot ++ (2 dishes !?)	medium constitutive promoter <-> ECFP	4+7
L110	a lot ++	medium constitutive promoter <-> gfp tripart	4+7
L111	a lot ++	weak constitutive promoter <-> ECFP	3+5
L112	a lot ++	weak constitutive promoter <-> gfp tripart	3+5
L113	1	Arac pBAD <-> ECFP	5+2.5
L114	2	Arac pBAD <-> gfp tripart	5+2.5
L115	a lot ++	pLas <-> ECFP	7+11
L116	a lot ++	pLas <-> gfp tripart	7+11
L117	+/- 30	Double terminator <-> YFP	4+3
L118	+/- 30	Double terminator <-> mRFP	4+2
L119	+/- 30	Double terminator <-> LasR activator (+LVA)	4+2
L120	a lot ++	tetR repressible promoter <-> ECFP	6+7.5
L121	a lot ++	tetR repressible promoter <-> gfp tripart	6+7.5
L122	0	B0034rbs-LasI <-> ECFP	7+3.5
L123	1	B0034rbs-LasI <-> gfp tripart	7+3.5
L124	a lot ++	tetR repble promoter <-> ECFP	6+7.5
L125	a lot ++	tetR repble promoter <-> ECFP	6+7.5
L126	a lot ++	tetR repble promoter <-> ECFP	6+7.5
L127	a lot ++	tetR repble promoter <-> ECFP	6+7.5

@@ Line 4: / Line 4: @@
 [[Image:RBS+.jpg|thumb|RBS +]]
 [[Image:RBS-.jpg|thumb|RBS -]]
-Creation of 3 oligonucleotides ([[team:Paris/Notebook/Oligo|O140, O141, O142]]) in order to create a particular plasmid
+Creation of 3 oligonucleotides ([[team:Paris/Notebook/Oligo|O140, O141, O142]]) in order to create two particular plasmids.
+Those plasmids are very useful vectors to amplify promoters and to measure their forces using Standard Promoter Units.
+The principle is easy to understand.
+Instead of using a big biobrick containing Promoter, RBS, GFP and Terminators, the Biobrick used for the test is only the Promoter. The standard GFP tripart (E0240) is between the biobrick site, it is just between two PstI restriction sites.
+This plasmid can be used to amplify a promoter because only the promoter is in the biobrick.