Difference between revisions of "Team:Munich/thisisatest.html"
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<td>Notes:</td> | <td>Notes:</td> | ||
<td>pkD3 contains resistance cassettes with FRT-sites for pRED engineering <br> | <td>pkD3 contains resistance cassettes with FRT-sites for pRED engineering <br> | ||
| − | + | Incubate pRED at 30°C because of temperature sensitive promoter | |
<br> | <br> | ||
pNPTS138-R6KT is for knock-ins via RecA Recombineering | pNPTS138-R6KT is for knock-ins via RecA Recombineering | ||
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<tr> | <tr> | ||
<td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></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 tried chemical transformation of NEB Turbo next.</td> |
</tr> | </tr> | ||
</table> | </table> | ||
| Line 46: | Line 46: | ||
<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><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></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 Dh5a as a cloning organism.</td> |
</tr> | </tr> | ||
</table> | </table> | ||
| Line 74: | Line 74: | ||
<tr> | <tr> | ||
<td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> | <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> | ||
| − | <td> | + | <td>No colonies</td> |
</tr> | </tr> | ||
</table> | </table> | ||
| Line 98: | Line 98: | ||
<tr> | <tr> | ||
<td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></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> | ||
Revision as of 19:14, 16 October 2018
Transforming E.Coli NEB Turbo to amplify plasmids for pRED/ET Engineering
2018/05/07| Participants: | Dominic Schwarz |
| Protocol: | Electrocompetent transformation |
| Notes: | pkD3 contains resistance cassettes with FRT-sites for pRED engineering Incubate pRED at 30°C because of temperature sensitive promoter pNPTS138-R6KT is for knock-ins via RecA Recombineering |
| Results: | No colonies. we suspected electroporation to be a problem and tried chemical transformation of 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 Dh5a as a cloning organism. |
Redo: Transforming E.Coli Dh5a to amplify plasmids for pRED/ET Engineering
2018/05/24| Participants: | Dominic Schwarz |
| Protocol: | Electrocompetent transformation |
| Notes: | inoculate pRED at 30°C because of temperature sensitive promoter, pKD3 contains resistance cassette flanked by FRT sites |
| Results: | No colonies |
Transforming E.Coli Dh5a 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: | Mini Prep |
| Notes: | because pRED/ET could not be transformed, we got readily transformed cells from PD Dr. Jürgen Lassak from the LMU |
| Results: | pRED/ET: 37,5 ng/µl pNPTS138-R6KT: 60ng/ul |