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Team:TU-Delft/Notebook/2013/08/09/
From 2013.igem.org
Notebook
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9th of August
Lab work
The pT7 promoter was still an issue as there was too much of degraded plasmid DNA.
We then found a another pT7 promoter from the kit in pSB1C3 backbone. The reason for the previous pT7 failure was that it was grown using kanamycin, as the IGEM website clearly said that it should be grown in ampicillin though they are in AK backbone to get high copy number.
The bacillus backbones where transformed, grown , prepped and digested. The digested plasmid was then ligated with the AIP sensor device.
The peptide oligos were treated with kinase and hybridised. The peptides were dissolved with the help of Martijn Pinkse of Peter's group. We are also planning to check it with MS.
The strains for MIC determination was restreaked to keep them fresh for carrying out the experiment.
