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Team:Groningen/Template/MODULE/Notebook/secretion/week8
From 2014.igem.org
September 15 - September 19
Obtaining the anti-Staphylococcus aureus system (p2s) and the anti-Pseudomonas aeruginosa system(pLASs)
Both systems were assembled by using Gibson assembly
Downstream of the pLas a RBS is attached together with the promoter P2 and dspB with tail PCR
A Gibson tail was attached with PCR to the P2 with RBS, dspB with RBS, aiiA and the double terminator
ssUSP45DspB, and nisA was added and incubated for 90 minutes at 50 °C
The Gibson fragment was amplified with gradient PCR
The construct was digested, checked on gel, and send for sequencing
Only 1200 base pairs of the construct could be found on gel
For the pLASs 70ng of pLAS, 400ng of dspB+RBS, 290ng of aiiA and 50ng of double terminator. Though after amplifying the fragment and loading it on gel, only a 1200bp band could be found.
