Team:Paris/August 1

From 2008.igem.org

(Difference between revisions)
(→Protocol)
(→Conditions)
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* Preparation of 50µl of minipreps in water.
* Preparation of 50µl of minipreps in water.
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=== List of Minipreps ==
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|align="center"|MP L102
|align="center"|MP L102
|align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
|align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
-
|align="center"|
+
|align="center"|Strong rbs - YFP<br>D129 (BV) -  D118 (BI)
-
Strong rbs - YFP
+
-
<br>D129 (BV) -  D118 (BI)
+
|-
|-
|align="center"|MP L103
|align="center"|MP L103
|align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
|align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
-
|align="center"|
+
|align="center"|Strong rbs - GFP<br>D129 (BV) -  D122 (BI)
-
Strong rbs - GFP
+
-
<br>D129 (BV) -  D122 (BI)
+
|-
|-
|align="center"|MP L104   
|align="center"|MP L104   
|align="center"|[[Image:Part_icon_rbs.png]][[Image:Icon_coding.png]]
|align="center"|[[Image:Part_icon_rbs.png]][[Image:Icon_coding.png]]
-
|align="center"|
+
|align="center"|Strong rbs - lasR activator<br>D129 (BV) -  D114 (BI)
-
Strong rbs - lasR activator
+
-
<br>D129 (BV) -  D114 (BI)
+
|-
|-
|align="center"|MP L105
|align="center"|MP L105
|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
-
|align="center"|
+
|align="center"|Strong promoter - ECFP<br>D123 (BV) - D130 (BI)
-
Strong promoter - ECFP
+
-
<br>D123 (BV) - D130 (BI)
+
|-
|-
|align="center"|MP L106
|align="center"|MP L106
|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]]
-
|align="center"|
+
|align="center"|Strong promoter - gfp Tripart<br>D123 (BV) - D131 (BI)
-
Strong promoter - gfp Tripart
+
-
<br>D123 (BV) - D131 (BI)
+
|-
|-
|align="center"|MP L107
|align="center"|MP L107
|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
-
|align="center"|
+
|align="center"|Strongest promoter - ECFP<br>D103 (BV) - D131 (BI)
-
Strongest promoter - ECFP
+
-
<br>D103 (BV) - D131 (BI)
+
|-
|-
|align="center"|MP L108
|align="center"|MP L108
|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]]
-
|align="center"|
+
|align="center"|Strong promoter -  gfp Tripart<br>D103 (BV) - D130 (BI)
-
Strong promoter -  gfp Tripart
+
-
<br>D103 (BV) - D130 (BI)
+
|-
|-
|align="center"|MP L110
|align="center"|MP L110
|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]]
-
|align="center"|
+
|align="center"|Medium promoter - gfp Tripart<br>D124 (BV) - D131 (BI)
-
Medium promoter - gfp Tripart
+
-
<br>D124 (BV) - D131 (BI)
+
|-
|-
|align="center"|MP L111
|align="center"|MP L111
|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
-
|align="center"|
+
|align="center"|Weak promoter - ECFP<br>D104 (BV) - D130 (BI)  
-
Weak promoter - ECFP
+
-
<br>D104 (BV) - D130 (BI)  
+
|-
|-
|align="center"|MP L112
|align="center"|MP L112
|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
-
|align="center"|Weak promoter - gfp
+
|align="center"|Weak promoter - gfp<br>D104 (BV) - D131 (BI)
-
<br>D104 (BV) - D131 (BI)
+
|-
|-
|align="center"|MP L115
|align="center"|MP 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]]
-
|align="center"|
+
|align="center"|pLas - ECFP<br>D105 (BV) - D130 (BI)
-
pLas - ECFP
+
-
<br>D105 (BV) - D130 (BI)
+
|-
|-
|align="center"|MP L116
|align="center"|MP 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]]
-
|align="center"|
+
|align="center"|pLas - gfp Tripart<br>D105 (BV) - D131 (BI)
-
pLas - gfp Tripart
+
-
<br>D105 (BV) - D131 (BI)
+
|-
|-
|align="center"|MP L117
|align="center"|MP 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]]
-
|align="center"|
+
|align="center"|Double Terminator - yfp<br>D125 (FV) - D117 (FI)
-
Double Terminator - yfp
+
-
<br>D125 (FV) - D117 (FI)
+
|-
|-
|align="center"|MP L118
|align="center"|MP 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]]
-
|align="center"|
+
|align="center"|Double Terminator - rfp<br>D125 (FV) - D121 (FI)
-
Double Terminator - rfp
+
-
<br>D125 (FV) - D121 (FI)
+
|-
|-
|align="center"|MP L119
|align="center"|MP 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]]
-
|align="center"|
+
|align="center"|Double Terminator - lasR activator + LVA<br>D125 (FV) - D113 (FI)
-
Double Terminator - lasR activator + LVA
+
-
<br>D125 (FV) - D113 (FI)
+
|-
|-
|align="center"|MP L121
|align="center"|MP 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]]
-
|align="center"|
+
|align="center"|Strong promoter - gfp tripart<br>D106 (BV) - D130 (BI)
-
Strong promoter - gfp tripart
+
-
<br>D106 (BV) - D130 (BI)
+
|-
|-
|align="center"|MP L124
|align="center"|MP 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]]
-
|align="center"|
+
|align="center"|Strongest RBS - rfp<br>D102 (BV) - D122 (BI)
-
Strongest RBS - rfp
+
-
<br>D102 (BV) - D122 (BI)
+
|-
|-
|align="center"|MP L125
|align="center"|MP 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]]
-
|align="center"|
+
|align="center"|Strongest RBS - yfp<br>D102 (BV) - D118 (BI)
-
Strongest RBS - yfp
+
-
<br>D102 (BV) - D118 (BI)
+
|}
|}
-
 
-
 
== Stock Glycerol ==
== Stock Glycerol ==

Revision as of 12:28, 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.


= List of Minipreps

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


=== List of Strain stocked

Strain Ligation Description
S123 L104 Strong rbs - lasR activator
D129 (BV) - D114 (BI) 		Part icon rbs.pngIcon coding.png
S124 L105 Strong promoter - ECFP
D123 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon reporter.png
S125 L106 Strong promoter - gfp Tripart
D123 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
S126 L107 Strongest promoter - ECFP
D103 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon reporter.png
S127 L108 Strong promoter - gfp Tripart
D103 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
S128 L110 Medium promoter - gfp Tripart
D124 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
S129 L111 Weak promoter - ECFP
D104 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon reporter.png
S130 L112 Weak promoter - gfp
D104 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon reporter.png
S131 L115 pLas - ECFP
D105 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon reporter.png
S132 L116 pLas - gfp Tripart
D105 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
S133 L117 yfp - Double Terminator
D125 (FV) - D117 (FI) 		Part icon reporter.pngPart icon terminator.pngPart icon terminator.png
S134 L118 rfp - Double Terminator
D125 (FV) - D121 (FI) 		Part icon reporter.pngPart icon terminator.pngPart icon terminator.png
S135 L119 lasR activator + LVA - Double Terminator
D125 (FV) - D113 (FI) 		Icon coding.pngPart icon terminator.pngPart icon terminator.png
S136 L121 Strong promoter - gfp tripart
D106 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
S137 L124 Strongest RBS - rfp
D102 (BV) - D122 (BI) 		Part icon rbs.pngPart icon reporter.png
S138 L125 Strongest RBS - yfp
D102 (BV) - D118 (BI) 		Part icon rbs.pngPart icon reporter.png

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 Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png RBS-TetR-ECFP
L101 Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png RBS-tetR-GFP tripart
L113 Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png pBAD-ECFP
L114 Part icon regulatory.pngPart icon reporter.png pBAD-GFP
L120 Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png ptet-ECFP
L122 Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png RBS-lasI-ECFP
L123 Part icon rbs.pngIcon coding.pngPart icon reporter.pngPart icon terminator.png RBS-lasI-GFP-ter
L126 Part icon rbs.pngIcon coding.png RBS-lasR-LVA
Retrieved from "http://2008.igem.org/Team:Paris/August_1"