Team:INSA-Lyon/Notebook

From 2014.igem.org

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<a href="https://2014.igem.org/Team:INSA-Lyon/Notebook" class="hu-icon"><li class="icon">NOTEBOOK</li></a>
<a href="https://2014.igem.org/Team:INSA-Lyon/Notebook" class="hu-icon"><li class="icon">NOTEBOOK</li></a>
<a href="https://2014.igem.org/Team:INSA-Lyon/Protocol" class="hu-icon"><li class="icon">PROTOCOLS</li></a>
<a href="https://2014.igem.org/Team:INSA-Lyon/Protocol" class="hu-icon"><li class="icon">PROTOCOLS</li></a>
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<a href="https://2014.igem.org/Team:INSA-Lyon/Parts" class="hu-icon"><li class="icon">PARTS</li></a>
 
<a href="https://2014.igem.org/Team:INSA-Lyon/DataPage" class="hu-icon"><li class="icon">DATA PAGE</li></a>
<a href="https://2014.igem.org/Team:INSA-Lyon/DataPage" class="hu-icon"><li class="icon">DATA PAGE</li></a>
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<p> New tests with UV light to kill bacteria : 5 LB plates were spotted after exposure of 1,3,5 and 7 min to UV and genomic DNA was extracted from 50µL of each culture by heating 5min at 100°C and centrifuging 5 min at 14000 rpm.</p>
<p> New tests with UV light to kill bacteria : 5 LB plates were spotted after exposure of 1,3,5 and 7 min to UV and genomic DNA was extracted from 50µL of each culture by heating 5min at 100°C and centrifuging 5 min at 14000 rpm.</p>
<p> Electrophoresis gel revealed no bands, more DNA had to be dropped off on gel.</p>
<p> Electrophoresis gel revealed no bands, more DNA had to be dropped off on gel.</p>
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<p> Characterization of Curli promoter : Miniprep of pIG57 for future digestions and ligations.</p>
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Characterization of Curli promoter : Miniprep of pIG57 for future digestions and ligations.</p>
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<p> Characterization of Curli promoter : digested pIG57 (E + X), ligated to PC750 (E + S) or PC70 (E + S) and then transformed in Kanamycin plates.</p>
<p> Characterization of Curli promoter : digested pIG57 (E + X), ligated to PC750 (E + S) or PC70 (E + S) and then transformed in Kanamycin plates.</p>
<p> Culture of s225, s226+His1, s226+His2, s228, s229 for epifluorescence observation and immunocytochemistry. Culture of the same strains in 96 wells plates for Immunocytochemistry and for ThioflavineS staining.</p>
<p> Culture of s225, s226+His1, s226+His2, s228, s229 for epifluorescence observation and immunocytochemistry. Culture of the same strains in 96 wells plates for Immunocytochemistry and for ThioflavineS staining.</p>
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<p>Ni-DMG test at normal pH</p>
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<p> Mesure of the fluorescence and OD of the 96 wells plate with ThS with the Tecan with or without biofilm resuspension.</p>
<p> Mesure of the fluorescence and OD of the 96 wells plate with ThS with the Tecan with or without biofilm resuspension.</p>
<p> Culture of s225 and s226His2.</p>
<p> Culture of s225 and s226His2.</p>
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<p> Culture of each strain for icp-ms nickel test pH</p>
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<p> Culture of s204, s227, His1, His2, WT and CsgB- pkkCsgD for MET observation.</p>
<p> Culture of s204, s227, His1, His2, WT and CsgB- pkkCsgD for MET observation.</p>
<p>Culture of s225, s226+His2 in 96 wells plate. </p>
<p>Culture of s225, s226+His2 in 96 wells plate. </p>
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<p> ICP-MS nickel test at 3 concentrations</p>
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<p> Immunolabeling with 2 antibodies and observation with the epifluorescence microscope.</p>
<p> Immunolabeling with 2 antibodies and observation with the epifluorescence microscope.</p>
<p> Red Congo test on DH5α and 326 in batch.</p>
<p> Red Congo test on DH5α and 326 in batch.</p>
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<p> <p>ICP-MS nickel results</p></p>
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           <ul id="contenu6" style="list-style-type: none !important;display:none;">
           <ul id="contenu6" style="list-style-type: none !important;display:none;">
               <li><p>
               <li><p>
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<h6>2/10 </h6>
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<p>
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Pre-cultures of the negative control, positive control, PHC46 (P70-GFP) and PHC47(P750-GFP).
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</p>
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<h6>3/10 </h6>
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<p>
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We distributed the blank, growth control, positive control for GFP expression, PHC46 (P70-GFP) and PHC47 (P750-GFP) in a 96 well plate and explored the GFP expression at 37 degrees.
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</p>
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<h6>3/10 </h6>
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<p>
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We gathered the results at 37°C and interpreted the results of GFP expression from the P70 or P750 long promoter upstream.
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</p>
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<h6>4/10 </h6>
<h6>4/10 </h6>
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<p> Petri dish nickel test on the Petri dishes containing the best clones for each one of the newly transformed strains 224, 266, 267.</p>
<p> Petri dish nickel test on the Petri dishes containing the best clones for each one of the newly transformed strains 224, 266, 267.</p>
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<p>
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Pre-cultures of the negative control, positive control, PHC46 (P70-GFP) and PHC47(P750-GFP).
 +
</p>
<h6>8/10  </h6>
<h6>8/10  </h6>
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<p>Cultured the best clones for each one of the newly transformed strains 224, 266, 267, 325 and 334 in a 24 wells plate. </p>
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<p>
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Cultured the best clones for each one of the newly transformed strains 224, 266, 267, 325 and 334 in a 24 wells plate.
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</p>
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<p>
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We distributed the blank, growth control, positive control for GFP expression, PHC46 (P70-GFP) and PHC47 (P750-GFP) in a 96 well plate and explored the GFP expression at 30 degrees.
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</p>
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<p> Red congo and adherence tests on the newly transformed strains 224, 266, 267, 325 and 334 cultured in the 24 wells plate.</p>
<p> Red congo and adherence tests on the newly transformed strains 224, 266, 267, 325 and 334 cultured in the 24 wells plate.</p>
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<p>
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We interpreted and compared the results of GFP expression from the P70 or P750 long promoter upstream.
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</p>
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Latest revision as of 00:55, 18 October 2014

Curly'on - IGEM 2014 INSA-LYON

Here you can find a summary of all the experiments we made throughout the project.

To download a pdf with plasmids' description, resistance and names click here

To download a pdf with the strains' description, resistance and names click here


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