Team:Paris/August 1

From 2008.igem.org

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Revision as of 12:22, 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
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