Difference between revisions of "Team:Munich/thisisatest.html"
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<td>Results:</td> | <td>Results:</td> | ||
| − | <td>pRED/ET: 37,5 ng/ | + | <td>pRED/ET: 37,5 ng/µl |
pNPTS138-R6KT: 60ng/ul | pNPTS138-R6KT: 60ng/ul | ||
</td> | </td> | ||
Revision as of 12:21, 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 |