BIOTEC Dresden/Notepad/31 August 2010

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Ligation of parts

Restriction digest of following plasmid parts were repeated under the same conditions.

4a, 6e, 9b, 14f, 20f, 21f

The digest was heat inactivated for 20 minutes and then run on an agarose gel for confirming the success of the digest. The digests were fine and then gel purified after which concentration was measured.

Fusion Protein

A colony PCR was performed to confirm the insertion of our fused insert into the plasmid pETMM43 or pETMMZZ. 8 clones from each plate were picked and PCR amplified. Analysis on an agarose gel revealed two positive clones for luxI-protein A as well as for the construct proteinA-luxI. From these two positive clones overnight cultures were prepared.

AHL Sensor

AHL assay with part 18, for 3 hours, using AHL concentrations of 0.01-2000 nM, starting with O.D of 4.9.


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