Team:Paris/August 1

From 2008.igem.org

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(→List of transformation)
(→Protocol)
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|align="center"|'''Strain'''
|align="center"|'''Strain'''
|align="center"|'''Ligation'''
|align="center"|'''Ligation'''
-
|colspan="2"|'''Description'''
+
|align="center"|'''Biobricks'''
 +
|align="center"|'''Description'''
|-   
|-   
-
|S123
+
|align="center"|S123
|align="center"|L104
|align="center"|L104
|align="center"|Strong rbs - lasR activator<br>D129 (BV) -  D114 (BI)   
|align="center"|Strong rbs - lasR activator<br>D129 (BV) -  D114 (BI)   
|[[Image:Part_icon_rbs.png]][[Image:Icon_coding.png]]
|[[Image:Part_icon_rbs.png]][[Image:Icon_coding.png]]
|-  
|-  
-
|S124
+
|align="center"|S124
|align="center"|L105
|align="center"|L105
|align="center"|Strong promoter - ECFP<br>D123 (BV) - D130 (BI)
|align="center"|Strong promoter - ECFP<br>D123 (BV) - D130 (BI)
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
|-  
|-  
-
|S125
+
|align="center"|S125
|align="center"|L106
|align="center"|L106
|align="center"|Strong promoter - gfp Tripart<br>D123 (BV) - D131 (BI)
|align="center"|Strong promoter - gfp Tripart<br>D123 (BV) - D131 (BI)
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
|-
|-
-
|S126
+
|align="center"|S126
|align="center"|L107
|align="center"|L107
|align="center"|Strongest promoter - ECFP<br>D103 (BV) - D131 (BI)
|align="center"|Strongest promoter - ECFP<br>D103 (BV) - D131 (BI)
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
|-   
|-   
-
|S127
+
|align="center"|S127
|align="center"|L108
|align="center"|L108
|align="center"|Strong promoter -  gfp Tripart<br>D103 (BV) - D130 (BI)
|align="center"|Strong promoter -  gfp Tripart<br>D103 (BV) - D130 (BI)
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
|-   
|-   
-
|S128
+
|align="center"|S128
|align="center"|L110
|align="center"|L110
|align="center"|Medium promoter - gfp Tripart<br>D124 (BV) - D131 (BI)
|align="center"|Medium promoter - gfp Tripart<br>D124 (BV) - D131 (BI)
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
|-  
|-  
-
|S129
+
|align="center"|S129
|align="center"|L111
|align="center"|L111
|align="center"|Weak promoter - ECFP<br>D104 (BV) - D130 (BI)  
|align="center"|Weak promoter - ECFP<br>D104 (BV) - D130 (BI)  
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
|-  
|-  
-
|S130
+
|align="center"|S130
|align="center"|L112
|align="center"|L112
|align="center"|Weak promoter - gfp<br>D104 (BV) - D131 (BI)
|align="center"|Weak promoter - gfp<br>D104 (BV) - D131 (BI)
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
|-  
|-  
-
|S131
+
|align="center"|S131
|align="center"|L115
|align="center"|L115
|align="center"|pLas - ECFP<br>D105 (BV) - D130 (BI)
|align="center"|pLas - ECFP<br>D105 (BV) - D130 (BI)
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
|-
|-
-
|S132
+
|align="center"|S132
|align="center"|L116
|align="center"|L116
|align="center"|pLas - gfp Tripart<br>D105 (BV) - D131 (BI)
|align="center"|pLas - gfp Tripart<br>D105 (BV) - D131 (BI)
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
|-   
|-   
-
|S133
+
|align="center"|S133
|align="center"|L117
|align="center"|L117
|align="center"|yfp - Double Terminator <br>D125 (FV) - D117 (FI)
|align="center"|yfp - Double Terminator <br>D125 (FV) - D117 (FI)
|[[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
|[[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
|-
|-
-
|S134
+
|align="center"|S134
|align="center"|L118
|align="center"|L118
|align="center"|rfp - Double Terminator<br>D125 (FV) - D121 (FI)
|align="center"|rfp - Double Terminator<br>D125 (FV) - D121 (FI)
|[[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
|[[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
|-
|-
-
|S135
+
|align="center"|S135
|align="center"|L119
|align="center"|L119
|align="center"|lasR activator + LVA - Double Terminator<br>D125 (FV) - D113 (FI)
|align="center"|lasR activator + LVA - Double Terminator<br>D125 (FV) - D113 (FI)
|[[Image:Icon_coding.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
|[[Image:Icon_coding.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
|-
|-
-
|S136
+
|align="center"|S136
|align="center"|L121
|align="center"|L121
|align="center"|Strong promoter - gfp tripart<br>D106 (BV) - D130 (BI)
|align="center"|Strong promoter - gfp tripart<br>D106 (BV) - D130 (BI)
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
|-
|-
-
|S137
+
|align="center"|S137
|align="center"|L124
|align="center"|L124
|align="center"|Strongest RBS - rfp<br>D102 (BV) - D122 (BI)
|align="center"|Strongest RBS - rfp<br>D102 (BV) - D122 (BI)
|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
|-
|-
-
|S138
+
|align="center"|S138
|align="center"|L125
|align="center"|L125
|align="center"|Strongest RBS - yfp<br>D102 (BV) - D118 (BI)
|align="center"|Strongest RBS - yfp<br>D102 (BV) - D118 (BI)

Revision as of 12:37, 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 reporter.pngPart icon terminator.pngPart icon terminator.png yfp - Double Terminator
D125 (FV) - D117 (FI)
MP L118 Part icon reporter.pngPart icon terminator.pngPart icon terminator.png rfp - Double Terminator
D125 (FV) - D121 (FI)
MP L119 Icon coding.pngPart icon terminator.pngPart icon terminator.png lasR activator + LVA - Double Terminator
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 Biobricks 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 RBS-TetR-ECFP Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
L101 RBS-tetR-GFP tripart Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
L113 pBAD-ECFP Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
L114 pBAD-GFP Part icon regulatory.pngPart icon reporter.png
L120 ptet-ECFP Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
L122 RBS-lasI-ECFP Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
L123 RBS-lasI-GFP-ter Part icon rbs.pngIcon coding.pngPart icon reporter.pngPart icon terminator.png
L126 RBS-lasR-LVA Part icon rbs.pngIcon coding.png
Retrieved from "http://2008.igem.org/Team:Paris/August_1"