Team:Paris/July 31

From 2008.igem.org

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(→Oligonucleotides design)
(→Transformation results)
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This plasmid can be used to amplify a promoter because only the promoter is in the biobrick.
This plasmid can be used to amplify a promoter because only the promoter is in the biobrick.
<br><br><br><br><br><br><br><br><br><br>
<br><br><br><br><br><br><br><br><br><br>
 +
 +
==Transformation results==
==Transformation results==
==Transformation results==
Line 18: Line 20:
|align="center"|'''Nb of colonies'''
|align="center"|'''Nb of colonies'''
|align="center"|'''Description'''
|align="center"|'''Description'''
-
|align="center"|'''Volume'''
+
|align="center"|'''Volume gel extraction'''
|-
|-
|align="center"|L100
|align="center"|L100
Line 142: Line 144:
|align="center"|L124
|align="center"|L124
|align="center"|a lot ++
|align="center"|a lot ++
-
|align="center"|tetR repble promoter <-> ECFP
+
|align="center"|strongest RBS (1) <-> RFP
-
|align="center"|6+7.5
+
|align="center"|6+5
|-
|-
|align="center"|L125
|align="center"|L125
|align="center"|a lot ++
|align="center"|a lot ++
-
|align="center"|tetR repble promoter <-> ECFP
+
|align="center"|strongest RBS (1) <-> YFP
-
|align="center"|6+7.5
+
|align="center"|6+10
|-
|-
|align="center"|L126
|align="center"|L126
-
|align="center"|a lot ++
+
|align="center"|no dish found!!!
-
|align="center"|tetR repble promoter <-> ECFP
+
|align="center"|strongest RBS (1) <-> activator (+LVA)
-
|align="center"|6+7.5
+
|align="center"|6+4
|-
|-
|align="center"|L127
|align="center"|L127
-
|align="center"|a lot ++
+
|align="center"|no dish found!!!
-
|align="center"|tetR repble promoter <-> ECFP
+
|align="center"|Double terminator <-> GFP
-
|align="center"|6+7.5
+
|align="center"|6+3
-
|
+
|-
 +
|align="center"|'''Control'''
 +
|align="center"|
 +
|align="center"|
 +
|align="center"|
 +
|-
 +
|align="center"|C1
 +
|align="center"|0
 +
|align="center"|D110
 +
|align="center"|3
 +
|-
 +
|align="center"|C2
 +
|align="center"|5
 +
|align="center"|D129
 +
|align="center"|5
 +
|-
 +
|align="center"|C3
 +
|align="center"|+/- 100
 +
|align="center"|D123
 +
|align="center"|5
 +
|-
 +
|align="center"|C4
 +
|align="center"|+/- 20
 +
|align="center"|D103
 +
|align="center"|5
 +
|-
 +
|align="center"|C5
 +
|align="center"|+/- 100
 +
|align="center"|Double terminator <-> GFP
 +
|align="center"|4
 +
|-
 +
|align="center"|C6
 +
|align="center"|+/- 100
 +
|align="center"|Double terminator <-> GFP
 +
|align="center"|3
 +
|-
 +
|align="center"|C7
 +
|align="center"|0
 +
|align="center"|Double terminator <-> GFP
 +
|align="center"|5
 +
|-
 +
|align="center"|C8
 +
|align="center"|+/- 20
 +
|align="center"|Double terminator <-> GFP
 +
|align="center"|7
 +
|-
 +
|align="center"|C9
 +
|align="center"|0
 +
|align="center"|Double terminator <-> GFP
 +
|align="center"|4
 +
|-
 +
|align="center"|C10
 +
|align="center"|0
 +
|align="center"|Double terminator <-> GFP
 +
|align="center"|6
 +
|-
 +
|align="center"|C11
 +
|align="center"|0
 +
|align="center"|Double terminator <-> GFP
 +
|align="center"|7
 +
|-
 +
|align="center"|C12
 +
|align="center"|+/- 10
 +
|align="center"|Double terminator <-> GFP
 +
|align="center"|6
 +
|-
 +
|align="center"|Positive control
 +
|align="center"|155 (transformation efficiency:1.5*10^7/ug)
 +
|align="center"|???
 +
|align="center"|
|}
|}

Revision as of 15:44, 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

Transformation results

Ligation name Nb of colonies Description Volume gel extraction
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 ++ strongest RBS (1) <-> RFP 6+5
L125 a lot ++ strongest RBS (1) <-> YFP 6+10
L126 no dish found!!! strongest RBS (1) <-> activator (+LVA) 6+4
L127 no dish found!!! Double terminator <-> GFP 6+3
Control
C1 0 D110 3
C2 5 D129 5
C3 +/- 100 D123 5
C4 +/- 20 D103 5
C5 +/- 100 Double terminator <-> GFP 4
C6 +/- 100 Double terminator <-> GFP 3
C7 0 Double terminator <-> GFP 5
C8 +/- 20 Double terminator <-> GFP 7
C9 0 Double terminator <-> GFP 4
C10 0 Double terminator <-> GFP 6
C11 0 Double terminator <-> GFP 7
C12 +/- 10 Double terminator <-> GFP 6
Positive control 155 (transformation efficiency:1.5*10^7/ug) ???
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