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Team:Amsterdam/8 September 2011
From 2011.igem.org
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08-09-2011
Constructs
The following parts were miniprepped:
- Cpn10
- Cpn60
- CspC
- K538100
- K538101
- K538104
- K538120
- K538130
- K538200
- Shednak
- R0010
- K206000
Digestion
Constructs making using the 2a assembly method. Digestion of different parts to construct the following assemblies:
- R0010+ B0034 + CspC
- R0010+ B0034 + Cpn60
- R0010+ B0034 + Shednak
- K206000+ B0034 + CspC
- K206000+ B0034 + Cpn60
- K206000+ B0034 + Shednak
These assemblies were ligated ON at 16°C.
Backbone transferring
Digestion of different parts to tranfer to the Chloramphenicol backbone:
- All genes
- The constructs we've miniprepped today
Gel purification
The different parts were put on gel and purified with the qiaquick gel extraction kit. Afterwards the concentration of the parts were checked on gel. The following parts for the new assembly showed a good result:
- K538130
- K206000
- K538101
K5389120 The following parts for the backbone transferring showed a good result:
- Cpn60
- K538200
- K538101
K538200 Checking
The assembly was checked on gel after digestion with:
- PvuI + PvuII
- PvuI + HindIII
On gel this gave the right band sizes.
