Team:Paris/August 1

From 2008.igem.org

(Difference between revisions)
(→Stock Glycerol)
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* 1mL culture in LB
* 1mL culture in LB
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 +
=== List of Strain stocked
{|border="1"  
{|border="1"  
-
|Strain
+
|align="center"|'''Strain'''
-
|Ligation
+
|align="center"|'''Ligation'''
-
|Description
+
|colspan="2"'''Description'''
|-   
|-   
|S123
|S123
|align="center"|L104
|align="center"|L104
-
|align="center"|[[Image:Part_icon_rbs.png]][[Image:Icon_coding.png]]
+
|align="center"|Strong rbs - lasR activator<br>D129 (BV) -  D114 (BI)   
-
Strong rbs - lasR activator
+
|[[Image:Part_icon_rbs.png]][[Image:Icon_coding.png]]
-
<br>D129 (BV) -  D114 (BI)   
+
|-  
|-  
|S124
|S124
|align="center"|L105
|align="center"|L105
-
|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
+
|align="center"|Strong promoter - ECFP<br>D123 (BV) - D130 (BI)
-
Strong promoter - ECFP
+
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
-
<br>D123 (BV) - D130 (BI)
+
|-  
|-  
|S125
|S125
|align="center"|L106
|align="center"|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"|Strong promoter - gfp Tripart<br>D123 (BV) - D131 (BI)
-
Strong promoter - gfp Tripart
+
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-
<br>D123 (BV) - D131 (BI)
+
|-
|-
|S126
|S126
|align="center"|L107
|align="center"|L107
-
|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
+
|align="center"|Strongest promoter - ECFP<br>D103 (BV) - D131 (BI)
-
Strongest promoter - ECFP
+
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
-
<br>D103 (BV) - D131 (BI)
+
|-   
|-   
|S127
|S127
|align="center"|L108
|align="center"|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"|Strong promoter -  gfp Tripart<br>D103 (BV) - D130 (BI)
-
Strong promoter -  gfp Tripart
+
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-
<br>D103 (BV) - D130 (BI)
+
|-   
|-   
|S128
|S128
|align="center"|L110
|align="center"|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"|Medium promoter - gfp Tripart<br>D124 (BV) - D131 (BI)
-
Medium promoter - gfp Tripart
+
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-
<br>D124 (BV) - D131 (BI)
+
|-  
|-  
|S129
|S129
|align="center"|L111
|align="center"|L111
-
|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
+
|align="center"|Weak promoter - ECFP<br>D104 (BV) - D130 (BI)
-
Weak promoter - ECFP
+
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
-
<br>D104 (BV) - D130 (BI)
+
|-  
|-  
|S130
|S130
|align="center"|L112
|align="center"|L112
-
|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
+
|align="center"|Weak promoter - gfp<br>D104 (BV) - D131 (BI)
-
Weak promoter - gfp
+
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
-
<br>D104 (BV) - D131 (BI)
+
|-  
|-  
|S131
|S131
|align="center"|L115
|align="center"|L115
-
|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
+
|align="center"|pLas - ECFP<br>D105 (BV) - D130 (BI)
-
pLas - ECFP
+
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
-
<br>D105 (BV) - D130 (BI)
+
|-
|-
|S132
|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"|pLas - gfp Tripart<br>D105 (BV) - D131 (BI)
-
pLas - gfp Tripart
+
[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-
<br>D105 (BV) - D131 (BI)
+
|-   
|-   
|S133
|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"|Double Terminator - yfp<br>D125 (FV) - D117 (FI)
-
Double Terminator - yfp
+
|[[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_reporter.png]]
-
<br>D125 (FV) - D117 (FI)
+
|-
|-
|S134
|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"|Double Terminator - rfp<br>D125 (FV) - D121 (FI)
-
Double Terminator - rfp
+
|[[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_reporter.png]]
-
<br>D125 (FV) - D121 (FI)
+
|-
|-
|S135
|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"|Double Terminator - lasR activator + LVA<br>D125 (FV) - D113 (FI)
-
Double Terminator - lasR activator + LVA
+
|[[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]][[Image:Icon_coding.png]]
-
<br>D125 (FV) - D113 (FI)
+
|-
|-
|S136
|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"|Strong promoter - gfp tripart<br>D106 (BV) - D130 (BI)
-
Strong promoter - gfp tripart
+
|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-
<br>D106 (BV) - D130 (BI)
+
|-
|-
|S137
|S137
|align="center"|L124
|align="center"|L124
-
|align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
+
|align="center"|Strongest RBS - rfp<br>D102 (BV) - D122 (BI)
-
Strongest RBS - rfp
+
|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
-
<br>D102 (BV) - D122 (BI)
+
|-
|-
|S138
|S138
|align="center"|L125
|align="center"|L125
-
|align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
+
|align="center"|Strongest RBS - yfp<br>D102 (BV) - D118 (BI)
-
Strongest RBS - yfp
+
|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
-
<br>D102 (BV) - D118 (BI)
+
|}
|}
-
 
-
 
== Transformations ==
== Transformations ==

Revision as of 12:20, 5 August 2008

← Yesterday

↓ Calendar ↑

Tomorrow →


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


=== List of Strain stocked

Strain Ligation colspan="2"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 Double Terminator - yfp
D125 (FV) - D117 (FI) 		Part icon terminator.pngPart icon terminator.pngPart icon reporter.png
S134 L118 Double Terminator - rfp
D125 (FV) - D121 (FI) 		Part icon terminator.pngPart icon terminator.pngPart icon reporter.png
S135 L119 Double Terminator - lasR activator + LVA
D125 (FV) - D113 (FI) 		Part icon terminator.pngPart icon terminator.pngIcon coding.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"