Difference between revisions of "Team:Munich/thisisatest.html"
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<div class="event-info"> | <div class="event-info"> | ||
| − | + | <h4>Transforming E.Coli NEB Turbo to Amplify Plasmids for pRED/ET Engineering</h4> | |
| − | <h4>Transforming E.Coli NEB Turbo to | + | <em>2018/05/07</em> |
<table class="table table-borderless"> | <table class="table table-borderless"> | ||
<tr> | <tr> | ||
<td>Participants:</td> | <td>Participants:</td> | ||
| − | <td>Dominic</td> | + | <td>Dominic Schwarz</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Protocol:</td> | <td>Protocol:</td> | ||
| − | <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> | ||
<tr> | <tr> | ||
<td>Notes:</td> | <td>Notes:</td> | ||
| − | <td>pkD3 contains resistance cassettes with FRT-sites for pRED engineering | + | <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. |
| − | + | ||
| − | + | ||
| − | pNPTS138-R6KT is for | + | |
</td> | </td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td>Results:</td> | + | <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> |
| − | <td> | + | <td>No colonies. We suspected electroporation to be a problem and switched to chemical transformation in NEB Turbo next.</td> |
</tr> | </tr> | ||
</table> | </table> | ||
| − | + | <h4>Redo: Transforming E.Coli NEB Turbo to Amplify Plasmids for pRED/ET Engineering</h4> | |
| − | + | <em>2018/05/18</em> | |
| − | <h4>Transforming E.Coli NEB Turbo to | + | |
<table class="table table-borderless"> | <table class="table table-borderless"> | ||
<tr> | <tr> | ||
<td>Participants:</td> | <td>Participants:</td> | ||
| − | <td>Dominic</td> | + | <td>Dominic Schwarz</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Protocol:</td> | <td>Protocol:</td> | ||
| − | <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> | ||
<tr> | <tr> | ||
<td>Notes:</td> | <td>Notes:</td> | ||
| − | <td> | + | <td>Inoculate pRED at 30°C because of temperature sensitive promoter.</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td>Results:</td> | + | <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> |
| − | <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> | |
| − | <h4>Transforming E.Coli | + | <em>2018/05/24</em> |
<table class="table table-borderless"> | <table class="table table-borderless"> | ||
<tr> | <tr> | ||
<td>Participants:</td> | <td>Participants:</td> | ||
| − | <td>Dominic</td> | + | <td>Dominic Schwarz</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Protocol:</td> | <td>Protocol:</td> | ||
| − | <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> | ||
<tr> | <tr> | ||
<td>Notes:</td> | <td>Notes:</td> | ||
| − | <td>inoculate pRED at 30°C because of temperature sensitive promoter, pKD3 contains resistance cassette flanked by FRT sites</td> | + | <td>inoculate pRED at 30°C because of temperature sensitive promoter, pKD3 contains resistance cassette flanked by FRT sites.</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td>Results:</td> | + | <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> |
| − | <td> | + | <td>We got cells containing the plasmid from PD Dr. Jürgen Lassak of the LMU.</td> |
</tr> | </tr> | ||
</table> | </table> | ||
| − | + | <h4>Transforming E.Coli Dh5α to Amplify Plasmids for pRED/ET Engineering</h4> | |
| − | + | <em>2018/05/25</em> | |
| − | <h4>Transforming E.Coli | + | |
<table class="table table-borderless"> | <table class="table table-borderless"> | ||
<tr> | <tr> | ||
<td>Participants:</td> | <td>Participants:</td> | ||
| − | <td>Dominic</td> | + | <td>Dominic Schwarz</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Protocol:</td> | <td>Protocol:</td> | ||
| − | <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> | ||
<tr> | <tr> | ||
<td>Notes:</td> | <td>Notes:</td> | ||
| − | <td>pKD3 contains resistance cassette flanked by FRT sites</td> | + | <td>pKD3 contains resistance cassette flanked by FRT sites.</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td>Results:</td> | + | <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> |
| − | <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> | </tr> | ||
</table> | </table> | ||
| − | + | <h4>DNA Preparation for pRED/ET Engineering</h4> | |
| − | <h4>DNA | + | <em>2018/05/26</em> |
<table class="table table-borderless"> | <table class="table table-borderless"> | ||
<tr> | <tr> | ||
<td>Participants:</td> | <td>Participants:</td> | ||
| − | <td>Dominic</td> | + | <td>Dominic Schwarz</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Protocol:</td> | <td>Protocol:</td> | ||
| − | <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> | ||
<tr> | <tr> | ||
<td>Notes:</td> | <td>Notes:</td> | ||
| − | <td> | + | <td>The cells from PD Dr. Jürgen Lassak were used for DNA preparation.</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td>Results:</td> | + | <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> |
| − | <td>pRED/ET: 37,5 ng/ | + | <td>Concentration of the plasmids obtains from Miniprep: pRED/ET: 37,5 ng/µl |
| − | pNPTS138-R6KT: 60ng/ | + | pNPTS138-R6KT: 60ng/µl |
</td> | </td> | ||
</tr> | </tr> | ||
| Line 121: | Line 127: | ||
</div> | </div> | ||
| + | </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 |