Team:Munich/thisisatest.html
Transforming E.Coli NEB Turbo to amplify plasmids for pRED/ET Engineering
2018/05/07| Participants: | Dominic |
| Protocol: | epo. Trafo |
| 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 |
| Protocol: | chem Trafo |
| 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 |
| Protocol: | epo. Trafo |
| 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 |
| Protocol: | chem. Trafo |
| 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 |
| 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/ul pNPTS138-R6KT: 60ng/ul |