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V08_13
V_08_13_1 Overnight cultures
- Experiment:
5 ml LB-medium was inoculated with the fast chemotaxis strains MG1655 and RP437, respectively. 5 ml LB-medium with ampicillin was inoculated with Δtar_18C_tar and Δtar_18C_rfp.
- Observations & Results:
tar strains were used in a new separation assay. The strains contained the plasmid J62001.
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V08_14
V_08_14_1 Separation assay
- Experiment:
Repetition of V_08_09_1. Used tryptone and AS-mix as attractants. Also MG1655 and RP437 were equally mixed with Δtar_18C_rfp according to the OD600 measurement.
- Observations & Results:
M9-plates with AS-mix as attractant:MG1655 and RP437 showed minor swimming. Δtar-strains did not swim. M9-plates with tryptone as attractant: MG1655 shows minor swimming, the other strains not. The swimming has no direction! There was no chemotaxis observed. RFP expression not visible! The swimming cells are either not red enough or do not express the RFP. But they still are resisitant against ampicillin.
V_08_14_2 Motility assay with group 2 constructs
- Experiment:
The motility assay was performed in cooperation with group 2. The experiment is explained in more detail in the group 2 notebook.
- Observations & Results:
The observations & results can be found in the group 2 notebook.
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V08_15
V_08_15_1 Chemical transformation of constitutive promoters and tar in the plasmid pSB1C3 in E .coli DH10B Δtar
- Experiment:
The promoter constructs with the quik-changed tar gene were ligated with the pSB1C3 plasmid by group 3. The transformation was performed with chemically competent E .coli DH10B Δtar according to the transformation protocol.
- Observations & Results:
There was a bacterial lawn on the trafo plates. In future transformations the trafo should be divided: 100 µl on one selection plate and the rest on another.
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V08_17
V_08_17_1 Selection of fast MG1655 strains
- Experiment:
MG1655 strains from plates of V_08_14 were cut out 2 times. The agar was incubated in 1 ml LB-medium at 37°C and 180 rpm for 1 hour. The cultures were spun down for 10 mins at 1,5k X g. The supernatant was discarded and the pellet was resuspended in the remaining LB-medium. Ca. 8 µl were dropped on fresh M9 plates with AS-mix in the Whatman Paper. Subsequently, the plates were incubated at 35°C.
- Observations & Results:
Swimming was observed and the swimming front was cut out and the same procedure was repeated.
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V08_19
V_08_19_1 Overnight cultures
- Experiment:
E. coli DH10B and BL21 with various group two constructs were used to inoculate 5 ml LB with the corresponding antibiotics. Cultures were grown over night at 37°C and 180 rpm.
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