Team:Paris/August 1

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Revision as of 12:48, 5 August 2008

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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	Strong rbs - YFP D129 (BV) - D118 (BI)
MP L103	Strong rbs - GFP D129 (BV) - D122 (BI)
MP L104	Strong rbs - lasR activator D129 (BV) - D114 (BI)
MP L105	Strong promoter - ECFP D123 (BV) - D130 (BI)
MP L106	Strong promoter - gfp Tripart D123 (BV) - D131 (BI)
MP L107	Strongest promoter - ECFP D103 (BV) - D131 (BI)
MP L108	Strong promoter - gfp Tripart D103 (BV) - D130 (BI)
MP L110	Medium promoter - gfp Tripart D124 (BV) - D131 (BI)
MP L111	Weak promoter - ECFP D104 (BV) - D130 (BI)
MP L112	Weak promoter - gfp D104 (BV) - D131 (BI)
MP L115	pLas - ECFP D105 (BV) - D130 (BI)
MP L116	pLas - gfp Tripart D105 (BV) - D131 (BI)
MP L117	yfp - Double Terminator D125 (FV) - D117 (FI)
MP L118	rfp - Double Terminator D125 (FV) - D121 (FI)
MP L119	lasR activator + LVA - Double Terminator D125 (FV) - D113 (FI)
MP L121	Strong promoter - gfp tripart D106 (BV) - D130 (BI)
MP L124	Strongest RBS - rfp D102 (BV) - D122 (BI)
MP L125	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)
S124	L105	Strong promoter - ECFP D123 (BV) - D130 (BI)
S125	L106	Strong promoter - gfp Tripart D123 (BV) - D131 (BI)
S126	L107	Strongest promoter - ECFP D103 (BV) - D131 (BI)
S127	L108	Strong promoter - gfp Tripart D103 (BV) - D130 (BI)
S128	L110	Medium promoter - gfp Tripart D124 (BV) - D131 (BI)
S129	L111	Weak promoter - ECFP D104 (BV) - D130 (BI)
S130	L112	Weak promoter - gfp D104 (BV) - D131 (BI)
S131	L115	pLas - ECFP D105 (BV) - D130 (BI)
S132	L116	pLas - gfp Tripart D105 (BV) - D131 (BI)
S133	L117	yfp - Double Terminator D125 (FV) - D117 (FI)
S134	L118	rfp - Double Terminator D125 (FV) - D121 (FI)
S135	L119	lasR activator + LVA - Double Terminator D125 (FV) - D113 (FI)
S136	L121	Strong promoter - gfp tripart D106 (BV) - D130 (BI)
S137	L124	Strongest RBS - rfp D102 (BV) - D122 (BI)
S138	L125	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	ptet-ECFP
L122	RBS-lasI-ECFP
L123	RBS-lasI-GFP-ter
L126	RBS-lasR-LVA

@@ Line 10: / Line 10: @@
-=== List of Minipreps ==
+=== List of Minipreps ===
@@ Line 19: / Line 19: @@
 |-
 |align="center"|MP L102
+|align="center"|Strong rbs - YFP<br>D129 (BV) -  D118 (BI)
 |align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
-|align="center"|Strong rbs - YFP<br>D129 (BV) -  D118 (BI)
 |-
 |align="center"|MP L103
+|align="center"|Strong rbs - GFP<br>D129 (BV) -  D122 (BI)
 |align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
-|align="center"|Strong rbs - GFP<br>D129 (BV) -  D122 (BI)
 |-
 |align="center"|MP L104
-|align="center"|[[Image:Part_icon_rbs.png]][[Image:Icon_coding.png]]
 |align="center"|Strong rbs - lasR activator<br>D129 (BV) -  D114 (BI)
+|align="center"|[[Image:Part_icon_rbs.png]][[Image:Icon_coding.png]]
 |-
 |align="center"|MP L105
-|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
 |align="center"|Strong promoter - ECFP<br>D123 (BV) - D130 (BI)
+|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
 |-
 |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"|Strong promoter - gfp Tripart<br>D123 (BV) - D131 (BI)
+|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
 |-
 |align="center"|MP L107
-|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
 |align="center"|Strongest promoter - ECFP<br>D103 (BV) - D131 (BI)
+|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
 |-
 |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"|Strong promoter -  gfp Tripart<br>D103 (BV) - D130 (BI)
+|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
 |-
 |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"|Medium promoter - gfp Tripart<br>D124 (BV) - D131 (BI)
+|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
 |-
 |align="center"|MP L111
+|align="center"|Weak promoter - ECFP<br>D104 (BV) - D130 (BI)
 |align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
-|align="center"|Weak promoter - ECFP<br>D104 (BV) - D130 (BI)
 |-
 |align="center"|MP L112
+|align="center"|Weak promoter - gfp<br>D104 (BV) - D131 (BI)
 |align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
-|align="center"|Weak promoter - gfp<br>D104 (BV) - D131 (BI)
 |-
 |align="center"|MP L115
+|align="center"|pLas - ECFP<br>D105 (BV) - D130 (BI)
 |align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
-|align="center"|pLas - ECFP<br>D105 (BV) - D130 (BI)
 |-
 |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"|pLas - gfp Tripart<br>D105 (BV) - D131 (BI)
+|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
 |-
 |align="center"|MP L117
-|align="center"|[[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
 |align="center"|yfp - Double Terminator<br>D125 (FV) - D117 (FI)
+|align="center"|[[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
 |-
 |align="center"|MP L118
-|align="center"|[[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
 |align="center"|rfp - Double Terminator<br>D125 (FV) - D121 (FI)
+|align="center"|[[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
 |-
 |align="center"|MP L119
-|align="center"|[[Image:Icon_coding.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
 |align="center"|lasR activator + LVA - Double Terminator<br>D125 (FV) - D113 (FI)
+|align="center"|[[Image:Icon_coding.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
 |-
 |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"|Strong promoter - gfp tripart<br>D106 (BV) - D130 (BI)
+|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
 |-
 |align="center"|MP L124
-|align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
 |align="center"|Strongest RBS - rfp<br>D102 (BV) - D122 (BI)
+|align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
 |-
 |align="center"|MP L125
-|align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
 |align="center"|Strongest RBS - yfp<br>D102 (BV) - D118 (BI)
+|align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
 |}

Team:Paris/August 1

From 2008.igem.org

Revision as of 12:48, 5 August 2008

Contents

MiniPreps

Conditions

List of Minipreps

Stock Glycerol

Protocol

Transformations

Protocol

List of transformation