Team:Paris/August 1

From 2008.igem.org

(Difference between revisions)
(→Protocol)
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Weak promoter - gfp
Weak promoter - gfp
<br>D104 (BV) - D131 (BI)
<br>D104 (BV) - D131 (BI)
-
|-
+
|-  
-
|- style="background: #dddddd;" 
+
|S131
-
| style="background: #D4E2EF;" |S131
+
|align="center"|L115
|align="center"|L115
|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
Line 193: Line 192:
<br>D105 (BV) - D130 (BI)
<br>D105 (BV) - D130 (BI)
|-
|-
-
|- style="background: #dddddd;" 
+
|S132
-
| style="background: #D4E2EF;" |S132
+
|align="center"|L116
|align="center"|L116
|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
pLas - gfp Tripart
pLas - gfp Tripart
<br>D105 (BV) - D131 (BI)
<br>D105 (BV) - D131 (BI)
-
|-
+
|-   
-
|- style="background: #dddddd;"  
+
|S133
-
| style="background: #D4E2EF;" |S133
+
|align="center"|L117
|align="center"|L117
|align="center"|[[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_reporter.png]]
|align="center"|[[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_reporter.png]]
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<br>D125 (FV) - D117 (FI)
<br>D125 (FV) - D117 (FI)
|-
|-
-
|- style="background: #dddddd;" 
+
|S134
-
| style="background: #D4E2EF;" |S134
+
|align="center"|L118
|align="center"|L118
|align="center"|[[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_reporter.png]]
|align="center"|[[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_reporter.png]]
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<br>D125 (FV) - D121 (FI)
<br>D125 (FV) - D121 (FI)
|-
|-
-
|- style="background: #dddddd;" 
+
|S135
-
| style="background: #D4E2EF;" |S135
+
|align="center"|L119
|align="center"|L119
|align="center"|[[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]][[Image:Icon_coding.png]]
|align="center"|[[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]][[Image:Icon_coding.png]]
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<br>D125 (FV) - D113 (FI)
<br>D125 (FV) - D113 (FI)
|-
|-
-
|- style="background: #dddddd;" 
+
|S136
-
| style="background: #D4E2EF;" |S136
+
|align="center"|L121
|align="center"|L121
|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
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<br>D106 (BV) - D130 (BI)
<br>D106 (BV) - D130 (BI)
|-
|-
-
|- style="background: #dddddd;" 
+
|S137
-
| style="background: #D4E2EF;" |S137
+
|align="center"|L124
|align="center"|L124
|align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
|align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
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<br>D102 (BV) - D122 (BI)
<br>D102 (BV) - D122 (BI)
|-
|-
-
|- style="background: #dddddd;" 
+
|S138
-
| style="background: #D4E2EF;" |S138
+
|align="center"|L125
|align="center"|L125
|align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
|align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]

Revision as of 11:10, 5 August 2008

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Contents

MiniPreps

Conditions

  • Use of Quiagen's protocol on all the clones cultivated on the 31th.
  • Preparation of 50µl of minipreps in water.


Name Description Biobricks
MP L102 Part icon rbs.pngPart icon reporter.png

Strong rbs - YFP
D129 (BV) - D118 (BI)

MP L103 Part icon rbs.pngPart icon reporter.png

Strong rbs - GFP
D129 (BV) - D122 (BI)

MP L104 Part icon rbs.pngIcon coding.png

Strong rbs - lasR activator
D129 (BV) - D114 (BI)

MP L105 Part icon regulatory.pngPart icon reporter.png

Strong promoter - ECFP
D123 (BV) - D130 (BI)

MP L106 Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png

Strong promoter - gfp Tripart
D123 (BV) - D131 (BI)

MP L107 Part icon regulatory.pngPart icon reporter.png

Strongest promoter - ECFP
D103 (BV) - D131 (BI)

MP L108 Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png

Strong promoter - gfp Tripart
D103 (BV) - D130 (BI)

MP L110 Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png

Medium promoter - gfp Tripart
D124 (BV) - D131 (BI)

MP L111 Part icon regulatory.pngPart icon reporter.png

Weak promoter - ECFP
D104 (BV) - D130 (BI)

MP L112 Part icon regulatory.pngPart icon reporter.png Weak promoter - gfp


D104 (BV) - D131 (BI)

MP L115 Part icon regulatory.pngPart icon reporter.png

pLas - ECFP
D105 (BV) - D130 (BI)

MP L116 Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png

pLas - gfp Tripart
D105 (BV) - D131 (BI)

MP L117 Part icon terminator.pngPart icon terminator.pngPart icon reporter.png

Double Terminator - yfp
D125 (FV) - D117 (FI)

MP L118 Part icon terminator.pngPart icon terminator.pngPart icon reporter.png

Double Terminator - rfp
D125 (FV) - D121 (FI)

MP L119 Part icon terminator.pngPart icon terminator.pngIcon coding.png

Double Terminator - lasR activator + LVA
D125 (FV) - D113 (FI)

MP L121 Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png

Strong promoter - gfp tripart
D106 (BV) - D130 (BI)

MP L124 Part icon rbs.pngPart icon reporter.png

Strongest RBS - rfp
D102 (BV) - D122 (BI)

MP L125 Part icon rbs.pngPart icon reporter.png

Strongest RBS - yfp
D102 (BV) - D118 (BI)


Stock Glycerol

Protocol

  • 1mL glycerol
  • 1mL culture in LB


Strain Ligation Description
S123 L104 Part icon rbs.pngIcon coding.png

Strong rbs - lasR activator
D129 (BV) - D114 (BI)

S124 L105 Part icon regulatory.pngPart icon reporter.png

Strong promoter - ECFP
D123 (BV) - D130 (BI)

S125 L106 Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png

Strong promoter - gfp Tripart
D123 (BV) - D131 (BI)

S126 L107 Part icon regulatory.pngPart icon reporter.png

Strongest promoter - ECFP
D103 (BV) - D131 (BI)

S127 L108 Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png

Strong promoter - gfp Tripart
D103 (BV) - D130 (BI)

S128 L110 Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png

Medium promoter - gfp Tripart
D124 (BV) - D131 (BI)

S129 L111 Part icon regulatory.pngPart icon reporter.png

Weak promoter - ECFP
D104 (BV) - D130 (BI)

S130 L112 Part icon regulatory.pngPart icon reporter.png

Weak promoter - gfp
D104 (BV) - D131 (BI)

S131 L115 Part icon regulatory.pngPart icon reporter.png

pLas - ECFP
D105 (BV) - D130 (BI)

S132 L116 Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png

pLas - gfp Tripart
D105 (BV) - D131 (BI)

S133 L117 Part icon terminator.pngPart icon terminator.pngPart icon reporter.png

Double Terminator - yfp
D125 (FV) - D117 (FI)

S134 L118 Part icon terminator.pngPart icon terminator.pngPart icon reporter.png

Double Terminator - rfp
D125 (FV) - D121 (FI)

S135 L119 Part icon terminator.pngPart icon terminator.pngIcon coding.png

Double Terminator - lasR activator + LVA
D125 (FV) - D113 (FI)

S136 L121 Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png

Strong promoter - gfp tripart
D106 (BV) - D130 (BI)

S137 L124 Part icon rbs.pngPart icon reporter.png

Strongest RBS - rfp
D102 (BV) - D122 (BI)

S138 L125 Part icon rbs.pngPart icon reporter.png

Strongest RBS - yfp
D102 (BV) - D118 (BI)

Transformations

Protocol

use of TOP10 Chemically competent cells

  • Defroze competent cells on ice during 5'
  • Add 5µl of DNA Ligation in 50µL of competent bacterias (or 1µL for the positive control puc19)
  • Incubate 30' on ice
  • Heat-shock the cells during 30" at 42°C without shaking
  • Put 2' on ice
  • Add 250µL of pre-warmed SOC medium (4°C)
  • Incubate 1h at 37°C under shaking (225rpm)
  • Spin at 5.000rpm during 30"
  • Remove 150µL of supernatant
  • Resuspent the pellet in the 150µL left
  • Spread on adequated plates
  • Incubate O/N at 37°C


List of transformation

Name Biobrick Description
L100 RBS-TetR-ECFP
L101 RBS-tetR-GFP tripart
L113 pBAD-ECFP
L114 pBAD-GFP
L120 tetR-ECFP
L122 RBS-lasI-ECFP
L123 RBS-lasI-GFP-ter
L126 RBS-lasR-LVA
Retrieved from "http://2008.igem.org/Team:Paris/August_1"