ULB/11 August 2009

From 2009.igem.org

(Difference between revisions)
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Start of construction (ligation) in Standard 10.
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1st building: the promoter (BBa_R0011) and RBS (BBa_B0034)in a plasmid destination which contain the brig CcdB (BBa_P1010) (it induces cell death) and a resistance gene. As our plasmids are resistant to ampicillin, the plasmid is chosen destination pSB1C3 with brick BBa_P1010( resistance to chloramphenicol).
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Début de construction (ligation) en Standard 10
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1er construction : le promoteur (BBa_R0011) et le RBS (BBa_B0034) le plasmide de destination doit contenir la brick ccdB (BBa_P1010) (induit la mort de la cellule) et un gène de résistance différent de celui des plasmides contenant le promoteur et le RBS. Comme nos plasmides sont résistants à l’ampicilline, on choisit le plasmide de destination pSB1C3 avec la brick BBa_P1010 celui-ci résistant au chloramphénicol.  
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===Laboratory manipulation===
===Laboratory manipulation===
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===Computer part===
===Computer part===
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* Commande des primers pour amplification de ccdA +son promoteur faible constitutif (mob promoteur, non issu d’E. coli)
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* Order of primers for amplification ccdA + its weak constitutive promoter (promoter mob, not from E. Coli)  
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* Création de BioBricks (ccdA dans le sens reverse et forward)
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Creating BioBricks (CCDA in the forward and reverse direction).
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*Creating BioBricks (CCDA in the forward and reverse direction)
** http://partsregistry.org/Part:BBa_K196000:Design
** http://partsregistry.org/Part:BBa_K196000:Design
** http://partsregistry.org/Part:BBa_K196001:Design
** http://partsregistry.org/Part:BBa_K196001:Design

Revision as of 17:07, 20 September 2009

Start of construction (ligation) in Standard 10.

1st building: the promoter (BBa_R0011) and RBS (BBa_B0034)in a plasmid destination which contain the brig CcdB (BBa_P1010) (it induces cell death) and a resistance gene. As our plasmids are resistant to ampicillin, the plasmid is chosen destination pSB1C3 with brick BBa_P1010( resistance to chloramphenicol).


Laboratory manipulation

  • Thermoporation de BBa_I51020 (plasmide de destination résistant à l’ampicilline) avec des bactéries thermocompétentes.
  • Digestion de BBa_I51020 avec les enzymes EcoRI et PstI

Computer part

  • Order of primers for amplification ccdA + its weak constitutive promoter (promoter mob, not from E. Coli)

Creating BioBricks (CCDA in the forward and reverse direction).

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