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Team:TU-Delft/Notebook/2013/08/18/
From 2013.igem.org
Notebook
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18th of August
Lab work
The cloning schedule was carried forward with the SUMO with the prohibited restriction sites. Some changes were made to clone SUMO that enables us to avoid the use EcoR I. The SUMO peptide cloning was done and we had some colonies in the plate. The colony PCR was performed but due to very high Tm of primers we did not get positive results. Again we are planning to do colony PCR by increasing the annealing temperatures of the primers.
The sequences for gene synthesis were made by removing the restriction sites without disturbing the amino acid sequence.
The sequencing results of bacillus AIP sensor was a failure. The sequencing showed that it had no inserts.
The ligation for pTet:cI was done and transformation was done on Sunday. The ligation for pBad:AgrAC:pP2 was done to aid one of the final constructs.
Some overlapping primers were made to splice His-SUMO-Peptide with TetR.
