Team:Paris/August 1

From 2008.igem.org

(Difference between revisions)
(→Protocol)
Line 11: Line 11:
=== List of Minipreps ===
=== List of Minipreps ===
-
 
{| border="1"  
{| border="1"  
Line 90: Line 89:
|align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
|align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
|}
|}
 +
 +
== Stock Glycerol ==
== Stock Glycerol ==
Line 186: Line 187:
|align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
|align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
|}
|}
 +
 +
== Transformations ==
== Transformations ==
-
 
=== Protocol ===
=== Protocol ===
-
 
+
''use of TOP10 Chemically competent cells''
-
use of TOP10 Chemically competent cells
+
* Defroze competent cells on ice during 5'
* Defroze competent cells on ice during 5'
Line 210: Line 211:
=== List of transformation ===
=== List of transformation ===
-
 
{| border="1"  
{| border="1"  

Revision as of 12:56, 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.


List of Minipreps

Name Biobricks Description
MP L102 Strong rbs - YFP
D129 (BV) - D118 (BI) 		Part icon rbs.pngPart icon reporter.png
MP L103 Strong rbs - GFP
D129 (BV) - D122 (BI) 		Part icon rbs.pngPart icon reporter.png
MP L104 Strong rbs - lasR activator
D129 (BV) - D114 (BI) 		Part icon rbs.pngIcon coding.png
MP L105 Strong promoter - ECFP
D123 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon reporter.png
MP L106 Strong promoter - gfp Tripart
D123 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
MP L107 Strongest promoter - ECFP
D103 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon reporter.png
MP L108 Strong promoter - gfp Tripart
D103 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
MP L110 Medium promoter - gfp Tripart
D124 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
MP L111 Weak promoter - ECFP
D104 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon reporter.png
MP L112 Weak promoter - gfp
D104 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon reporter.png
MP L115 pLas - ECFP
D105 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon reporter.png
MP L116 pLas - gfp Tripart
D105 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
MP L117 yfp - Double Terminator
D125 (FV) - D117 (FI) 		Part icon reporter.pngPart icon terminator.pngPart icon terminator.png
MP L118 rfp - Double Terminator
D125 (FV) - D121 (FI) 		Part icon reporter.pngPart icon terminator.pngPart icon terminator.png
MP L119 lasR activator + LVA - Double Terminator
D125 (FV) - D113 (FI) 		Icon coding.pngPart icon terminator.pngPart icon terminator.png
MP L121 Strong promoter - gfp tripart
D106 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
MP L124 Strongest RBS - rfp
D102 (BV) - D122 (BI) 		Part icon rbs.pngPart icon reporter.png
MP L125 Strongest RBS - yfp
D102 (BV) - D118 (BI) 		Part icon rbs.pngPart icon reporter.png


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 Description Biobrick
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"