Team:Warsaw/Calendar-Main/6 September 2009
From 2009.igem.org
Assembly of endosomal detection operon
Marcin
Task:1
- Transformation of chemocompetent E. coli TOP10 strain
Constructs to transform:
Methods:
- Detailed protocol of transformation is described here
Task 2:
- Digest isolated plasmid with Bax CDS to verify the correctness of the ligation:
Methods:
- Reaction mixture composition:
2 μl purified plasmid DNA product 1 μl RsaI (NEB) 2 μl Buffer Tango (Fermentas) 15 μl MQ water
- Digest was performed 3 hours
Results:
Comments:
- The enzym must have been deactivated due to presence of two forms of the plasmid. If circular plasmid is cut there are only one linear form in the sample. Althought in each case there was two forms - circular and supercoiled.
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