Team:Groningen/Template/MODULE/Notebook/secretion/week8

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For the pLASs 70ng of <i>pLAS</i>, 400ng of <i>dspB+RBS</i>, 290ng of <i>aiiA</i> and 50ng of double terminator. Though after amplifying the fragment and loading it on gel, only a 1200bp band could be found.
 
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Latest revision as of 00:35, 18 October 2014

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
 
 
 
 
 

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