Team:NTNU-Trondheim/Notebook/October

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<div id="top-section3"><center><img src="https://static.igem.org/mediawiki/2013/2/21/Logo2_NTNU.png" alt="header" border="0"  usemap="#igemmap" width="1000" height="400"></center></div>
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       <ul>
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         <li><a href='https://2013.igem.org/Team:NTNU-Trondheim/Project'><span>Project description</span></a></li>
         <li><a href='https://2013.igem.org/Team:NTNU-Trondheim/Project'><span>Project description</span></a></li>
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        <li><a href='https://2013.igem.org/Team:NTNU-Trondheim/novelapproach'><span>A novel approach</span></a></li>
         <li><a href='https://2013.igem.org/Team:NTNU-Trondheim/Model'><span>Modelling</span></a></li>
         <li><a href='https://2013.igem.org/Team:NTNU-Trondheim/Model'><span>Modelling</span></a></li>
         <li><a href='https://2013.igem.org/Team:NTNU-Trondheim/Experiments_and_Results'><span>Experiments and Results</span></a></li>
         <li><a href='https://2013.igem.org/Team:NTNU-Trondheim/Experiments_and_Results'><span>Experiments and Results</span></a></li>
         <li><a href='https://2013.igem.org/Team:NTNU-Trondheim/Parts'><span>BioBrick Parts</span></a></li>
         <li><a href='https://2013.igem.org/Team:NTNU-Trondheim/Parts'><span>BioBrick Parts</span></a></li>
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         <li class='last'><a href='https://2013.igem.org/Team:NTNU_Trondheim/Attributions'><span>Attributions</span></a></li>
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         <li class='last'><a href='https://2013.igem.org/Team:NTNU-Trondheim/Acknowledgements'><span>Acknowledgements</span></a></li>
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     <li><a href='https://2013.igem.org/Team:NTNU-Trondheim/Achievements'><span>Achievements</span></a></li>
     <li><a href='https://2013.igem.org/Team:NTNU-Trondheim/Achievements'><span>Achievements</span></a></li>
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     <li class='last'><a href='https://2013.igem.org/Team:NTNU-Trondheim/Medalcriteria'><span>Criteria</span></a></li>
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     <li class='last'><a href='https://2013.igem.org/Team:NTNU-Trondheim/Medalcriteria'><span>Medal criteria</span></a></li>
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     </ul>
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<p style="text-align:center; color:black; "> Wednesday 02 - Friday 04.10.2013</p> </div>
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As the first attempt to send in the tat_GFP_RFP construct to iGEM Headquarters failed, we redid the cloning as we did 26-28.09.2013 and sent in a new construct.
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Latest revision as of 22:43, 4 October 2013

Trondheim iGEM 2013

header
Mercury
October

Wednesday 02.10.2013

SDS-PAGE of bacterial samples with the tat_GFP_RFP (ER1) and tat_ProteinG construct



In order to determine if Protein G is produced in the S3-2B sample and wheater a GFP-RFP dimer is produced in the ER1 sample, a SDS-PAGE was performed. Liquid cell culture with ER1 (tat_GFP_RFP construct), S3-2B (tat_ProteinG construct) and wildtype ER2566 ''E.coli'' cells was centrifuged and the pallet was mixed with SDS loading buffer. This mix was then heated at 95 °C for 15 minutes. The SDS-PAGE results can be viewed in the figure below

Figure 1: SDS-PAGE of bacterial samples. From the left: wild type ER2566, tat_GFP_RFP and tat_ProteinG

There is evidently produced Protein G in the S3-2B sample as there is an additional band at the expected size of about 60 kDa. There is also an additional band at the ER1 sample of about 28-30 kDa. This half the expected size of about 60 kDa. This indicates that only RFP is produced in the bacterias.

SDS-PAGE of bacterial samples with the tat_GFP_RFP (ER1) and tat_ProteinG construct



A new test on the Pm/XylS promoter was performed with the same conditions as earlier.

Wednesday 02 - Friday 04.10.2013

As the first attempt to send in the tat_GFP_RFP construct to iGEM Headquarters failed, we redid the cloning as we did 26-28.09.2013 and sent in a new construct.