Difference between revisions of "Team:Warwick/Description"

 
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<h4>Characterisation of an existing part</h4>
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<h4>Characterisation of An Existing Part</h4>
 
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<p>As our project had a large focus on fluorescence, we decided to characterise a part from the existing iGEM registry that produces red fluorescent protein in order to determine whether we could use it in our project. We decided against using RFP, however here are the results and methods used in the characterisation.</p>
 
<p>As our project had a large focus on fluorescence, we decided to characterise a part from the existing iGEM registry that produces red fluorescent protein in order to determine whether we could use it in our project. We decided against using RFP, however here are the results and methods used in the characterisation.</p>
  
 
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<ul> <li> The JO4450 gene from the registry was cloned into three different plasmids (pSB1K3, pSB3K3, and pSB4K4) each with a different copy number. </li>
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<ul> <li> The J04450 gene from the registry was cloned into three different plasmids (pSB1K3, pSB3K3, and pSB4K4) each with a different copy number. </li>
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<li>Each plasmid was transformed electrocompetent into MG1655-Z1</li>
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<li>Each plasmid was transformed into electrocompetent MG1655-Z1</li>
 
<li>Three colonies of each plasmid were then inoculated in LB and grown overnight.</li>
 
<li>Three colonies of each plasmid were then inoculated in LB and grown overnight.</li>
 
<li>In the morning, each inoculation was refreshed in M9 + casein amino acid (500x dilution), induced with different concentrations of IPTG (0uM, 250uM, and 500uM IPTG) and grown for another 6 hours.</li>
 
<li>In the morning, each inoculation was refreshed in M9 + casein amino acid (500x dilution), induced with different concentrations of IPTG (0uM, 250uM, and 500uM IPTG) and grown for another 6 hours.</li>
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<li> They were then transferred into a 96 well plate (200uL per well) and had their fluorescence measured every 15 minutes.</li>
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<li> They were then transferred into a 96 well plate (200uL per well) and measured their fluorescence every 15 minutes.</li>
 
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<p>The results for our characterisation can be found on the JO4450 part page in the registry.</p>
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<p>The results for our characterisation can be found on the <a href="http://parts.igem.org/Part:BBa_J04450">J04450 part page in the registry</a>.
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Latest revision as of 19:15, 18 September 2015

Warwick iGEM 2015

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