Team:Groningen/Template/MODULE/Notebook/secretion/week3

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6 - 8 of August
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August 6 - August 8
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<b>Goal:</b> obtain all the biobricks necessary for the secreting systems of <i>P. aeruginosa</i> and <i>S. aureus</i>.
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Obtaining all the BioBricks needed for the secretion system to destroy <i>P. aeruginosa</i> and <i>S. aureus</i>
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<i>E. coli</i> was chemically transformed with 3 biobricks  in order to obtain them:
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<div class="item">BBa_I746104: P2-promotor</div>
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<div class="item">Performing three transformation to <i>E. coli</i>, one with promoter P2 (<a href="http://parts.igem.org/Part: BBa_I746104">BBa_I746104</a>), one with <i>lasI</i> (<a href="http://parts.igem.org/Part:BBa_K081009">BBa_K081009</a>), and one with <i>aiiA</i> (<a href="http://parts.igem.org/Part:BBa_C0060">BBa_C0060</a>)</div>
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<div class="item">BBa_K081009: LasI (for the Detection part)</div>
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<div class="item">The tranformated <i>E. coli</i> was divided in several concentrations over LB agar plates with Chloramphenicol</div>
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<div class="item">BBa_C0060: Aiia</div>
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<div class="item">Three glycerol stocks of <i>E. coli</i> DH5α containing the RBS (<a href="http://parts.igem.org/Part:BBa_ B0034">BBa_ B0034</a>), the double terminator (<a href="http://parts.igem.org/Part:BBa_ B0015">BBa_ B0015</a>) and the promoter LasR (<a href="http://parts.igem.org/Part:BBa_ R0079">BBa_ R0079</a>), were inoculated in LB medium, and grown overnight</div>
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<div class="item">The plasmids were isolated out of the overnight cultures, and checked on gel</div>
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After which 1, 10, 20, 50% was inoculated on Chloramphenicol LB-agar plates.
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The RBS, Double Terminators, and the promotor LAS Biobricks were already transformed and isolated.
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The clones were inoculated in liquid media were they would be prepared for mini-prep.
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The O/N culture was mini-prepped and checked on gel.
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Latest revision as of 22:06, 17 October 2014

August 6 - August 8
 
Obtaining all the BioBricks needed for the secretion system to destroy P. aeruginosa and S. aureus
 
Performing three transformation to E. coli, one with promoter P2 (BBa_I746104), one with lasI (BBa_K081009), and one with aiiA (BBa_C0060)
The tranformated E. coli was divided in several concentrations over LB agar plates with Chloramphenicol
Three glycerol stocks of E. coli DH5α containing the RBS (BBa_ B0034), the double terminator (BBa_ B0015) and the promoter LasR (BBa_ R0079), were inoculated in LB medium, and grown overnight
The plasmids were isolated out of the overnight cultures, and checked on gel
 
 
 
 
 

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