Lab notebooks
February - March
- Project selection
- Scientific discussuions
- Initial planning
April
- Lab selection
- Cloning scheme design
- Ordering of the compounds, oligos, PCR primers and other DNA fragments
May
- Introduction to the lab
- PCR amplification of required fragments
- Cloning of the level 0 plasmids
- Editing of the backbones available in the lab (e.g.inversions)
- Target selection
June
- Cloning most level 1 plasmids
- Cloning optimisation for golden gate
- Creation of promoter and RBS libraries for the genes required for the plasmid based EcORep system
- Design of the fluorescence based bridge recombinase efficiency assay
- Design of the DML library of IS622 variants
July
- Deletion of the csg operon in Marionette-Clo E. Coli strain to obtain SEM6 E.Coli, a strain that will be used for further transformations in course of the project
- Fluorescence based efficiency assay (GFP-mScarlet inversion, GFP insertion) to determine the initial efficiency of bridge recombinases towards selected target and donor sequences
August
- DML variant preparation
- Illumina sequencing
- Preliminary design of the selection plasmid antibiotic-based functionality assay
- Phage cloning
September
- Plating assay to determine the functionality of the designed selection plasmid
- Report preparation
- Phage propagation assay
October
- Presentation preparation
- Team Wiki creation
- Poster design