ULB/11 August 2009
From 2009.igem.org
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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 of BBa_I51020 (plasmid destination resistant to ampicillin) with thermo-skill bacteria thermocompétentes.
- Digestion of BBa_I51020 with EcoRI and 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).
- Creating BioBricks (CCDA in the forward and reverse direction)