Difference between revisions of "Team:UCL/Experiments"

 
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<h5>Experiments &amp; Protocols</h5>
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<p>Describe the experiments, research and protocols you used in your iGEM project.</p>
 
 
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<li> Protocols </li>
 
<li> Experiments </li>
 
<li>Documentation of the development of your project </li>
 
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<div id="transformation"><h2>Transformation protocol </h2>
 
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<li>(If using part from the distribution: resuspend the DNA in 10 ul of MiliQ water, making sure that it turns red. Wait 10 minutes before adding the DNA to cells)</li>
 
<li>Put a tube of NEB DH 5 alpha <i>E. coli</i> cells on ice and wait until they thaw completely. Divide the cells into 50 ul aliquotes. </li>
 
<li>Add 1 ul of plasmid DNA to 50 ul of cells. </li>
 
<li>Mix by carefully flicking the tube. Do not vortex or pipette in and out! </li>
 
<li>Place the mixture on ice for 30 minutes. </li>
 
<li>Heat shock the cells at 42 °C for 30 seconds and immediately put on back on ice.</li>
 
<li>Keep cells on ice for next 5 minutes. Do not mix. </li>
 
<li>Pipette 950 ul of SOC (or LB) media kept at room temperature into the mixture. <br/>
 
<li>Incubate the mixture at 37 °C for 60 minutes </li>
 
<li>Plate 50 ul of cells to the plate with appropriate antibiotic and incubate overnight at 37 °C</li>
 
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Latest revision as of 00:29, 6 August 2015