Team:Groningen/5 July 2010

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iGEM Groningen 2010

Hydrophobofilm
pushing coatings into a greener future

Week 27, Arend Jan


The pMASK-EH construct is cut with EcoRI and SpeI to get the fragments containing the ChpE and ChpH genes and ligate them in to the expression plasmid. A mix of both genes will be used for ligation and the clones will be checked afterwards for which gene they contain. 


- 10ul plasmid pNZ8901-bbs
- 1.5ul buffer Tango
- 0.5ul SpeI
- 3ul MQ
- perform digestion
- add 1.88ul buffer Tango
- add 0.5ul EcoRI

- 17.5ul plasmid pMASK-EH
- 2ul buffer Tango
- 0.5ul SpeI
- perform digestion
- add 2.5ul buffer Tango
- add 0.5ul EcoRI


Mix of fragments containing ChpE and ChpH genes at ~300bp
Restriction of expression backbone with EcoRI and SpeI



Tried to cut out the insertion from pNZ8901 and replace it with a part from a Plasmid with an identical backbone, all attempts failed however.




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