Difference between revisions of "Team:Munich/thisisatest.html"

 
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<div class="event-info">
 
<div class="event-info">
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<div>
 
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<h3>Transforming E.Coli NEB Turbo to amplify plasmids for pRED/ET Engineering</h3>
 
  
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Participants: Dominic
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<h4>Transforming E.Coli NEB Turbo to Amplify Plasmids for pRED/ET Engineering</h4>
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Protocol: epo. Trafo
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<em>2018/05/07</em>
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Notes: inoculate pRED at 30°C because of temperature sensitive promoter
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<table class="table table-borderless">
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Results: no colonies
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    <tr>
 +
 
 +
      <td>Participants:</td>
 +
      <td>Dominic Schwarz</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Protocol:</td>
 +
      <td>
 +
<a href="https://static.igem.org/mediawiki/2018/c/ca/T--Munich--WL1_Electrocompetent_transformation.pdf" target="_blank">Electro-transformation</a>
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</td>
 +
    </tr>
 +
    <tr>
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      <td>Notes:</td>
 +
      <td>pkD3 contains resistance cassettes with FRT-sites for pRED engineering. Due to the temperature sensitive promotor pNPTS138-R6KT which is used for bone-knockin via RecA recombineering, the cells were incubated at 30°C overnight.
 +
 
 +
</td>
 +
    </tr>
 +
<tr>
 +
      <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td>
 +
      <td>No colonies. We suspected electroporation to be a problem and switched to chemical transformation in NEB Turbo next.</td>
 +
    </tr>
 +
</table>
  
 +
<h4>Redo: Transforming E.Coli NEB Turbo to Amplify Plasmids for pRED/ET Engineering</h4>
 +
<em>2018/05/18</em>
 
<table class="table table-borderless">
 
<table class="table table-borderless">
 
     <tr>
 
     <tr>
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       <td>Mark</td>
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       <td>Otto</td>
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       <td>Participants:</td>
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       <td>Dominic Schwarz</td>
 
     </tr>
 
     </tr>
 
     <tr>
 
     <tr>
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       <td>Thornton</td>
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       <td>Protocol:</td>
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       <td>@fat</td>
+
       <td>
 +
<a href="https://static.igem.org/mediawiki/2018/a/a5/T--Munich--WL1_Chemical_transformation.pdf" target="_blank">Chemical Transformation</a>
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</td>
 
     </tr>
 
     </tr>
 
     <tr>
 
     <tr>
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       <td>the Bird</td>
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       <td>Notes:</td>
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       <td>@twitter</td>
+
       <td>Inoculate pRED at 30°C because of temperature sensitive promoter.</td>
 +
    </tr>
 +
<tr>
 +
      <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td>
 +
      <td>No colonies. We suspected the E. Coli NEB Turbo to be a problem and switched to E. Coli Dh5α.</td>
 
     </tr>
 
     </tr>
 
</table>
 
</table>
  
 +
<h4>Redo: Transforming E.Coli Dh5α to Amplify Plasmids for pRED/ET Engineering</h4>
 +
<em>2018/05/24</em>
 +
<table class="table table-borderless">
 +
    <tr>
  
 +
      <td>Participants:</td>
 +
      <td>Dominic Schwarz</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Protocol:</td>
 +
      <td>
 +
<a href="https://static.igem.org/mediawiki/2018/c/ca/T--Munich--WL1_Electrocompetent_transformation.pdf" target="_blank">Electro-transformation</a>
 +
</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Notes:</td>
 +
      <td>inoculate pRED at 30°C because of temperature sensitive promoter, pKD3 contains resistance cassette flanked by FRT sites.</td>
 +
    </tr>
 +
<tr>
 +
      <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td>
 +
      <td>We got cells containing the plasmid from PD Dr. Jürgen Lassak of the LMU.</td>
 +
    </tr>
 +
</table>
  
 +
<h4>Transforming E.Coli Dh5α to Amplify Plasmids for pRED/ET Engineering</h4>
 +
<em>2018/05/25</em>
 +
<table class="table table-borderless">
 +
    <tr>
  
 +
      <td>Participants:</td>
 +
      <td>Dominic Schwarz</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Protocol:</td>
 +
      <td>
 +
<a href="https://static.igem.org/mediawiki/2018/a/a5/T--Munich--WL1_Chemical_transformation.pdf" target="_blank">Chemical transformation</a>
 +
</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Notes:</td>
 +
      <td>pKD3 contains resistance cassette flanked by FRT sites.</td>
 +
    </tr>
 +
<tr>
 +
      <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td>
 +
      <td>No colonies. Because pRED/ET could not be transformed, we got readily transformed cells from PD Dr. Jürgen Lassak from the LMU.</td>
 +
    </tr>
 +
</table>
 +
 +
<h4>DNA Preparation for pRED/ET Engineering</h4>
 +
<em>2018/05/26</em>
 +
<table class="table table-borderless">
 +
    <tr>
 +
 +
      <td>Participants:</td>
 +
      <td>Dominic Schwarz</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Protocol:</td>
 +
      <td>
 +
<a href="https://international.neb.com/protocols/2015/11/20/monarch-plasmid-dna-miniprep-kit-protocol-t1010" target="_blank">Miniprep</a>
 +
</td>
 +
    </tr>
 +
    <tr>
 +
      <td>Notes:</td>
 +
      <td>The cells from PD Dr. Jürgen Lassak were used for DNA preparation.</td>
 +
    </tr>
 +
<tr>
 +
      <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td>
 +
      <td>Concentration of the plasmids obtains from Miniprep: pRED/ET: 37,5 ng/µl
 +
pNPTS138-R6KT: 60ng/µl
 +
</td>
 +
    </tr>
 +
</table>
  
 
</div>
 
</div>
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</div>
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</html>

Latest revision as of 00:11, 18 October 2018

Transforming E.Coli NEB Turbo to Amplify Plasmids for pRED/ET Engineering

2018/05/07
Participants: Dominic Schwarz
Protocol: Electro-transformation
Notes: pkD3 contains resistance cassettes with FRT-sites for pRED engineering. Due to the temperature sensitive promotor pNPTS138-R6KT which is used for bone-knockin via RecA recombineering, the cells were incubated at 30°C overnight.
Results: No colonies. We suspected electroporation to be a problem and switched to chemical transformation in NEB Turbo next.

Redo: Transforming E.Coli NEB Turbo to Amplify Plasmids for pRED/ET Engineering

2018/05/18
Participants: Dominic Schwarz
Protocol: Chemical Transformation
Notes: Inoculate pRED at 30°C because of temperature sensitive promoter.
Results: No colonies. We suspected the E. Coli NEB Turbo to be a problem and switched to E. Coli Dh5α.

Redo: Transforming E.Coli Dh5α to Amplify Plasmids for pRED/ET Engineering

2018/05/24
Participants: Dominic Schwarz
Protocol: Electro-transformation
Notes: inoculate pRED at 30°C because of temperature sensitive promoter, pKD3 contains resistance cassette flanked by FRT sites.
Results: We got cells containing the plasmid from PD Dr. Jürgen Lassak of the LMU.

Transforming E.Coli Dh5α to Amplify Plasmids for pRED/ET Engineering

2018/05/25
Participants: Dominic Schwarz
Protocol: Chemical transformation
Notes: pKD3 contains resistance cassette flanked by FRT sites.
Results: No colonies. Because pRED/ET could not be transformed, we got readily transformed cells from PD Dr. Jürgen Lassak from the LMU.

DNA Preparation for pRED/ET Engineering

2018/05/26
Participants: Dominic Schwarz
Protocol: Miniprep
Notes: The cells from PD Dr. Jürgen Lassak were used for DNA preparation.
Results: Concentration of the plasmids obtains from Miniprep: pRED/ET: 37,5 ng/µl pNPTS138-R6KT: 60ng/µl