June
18/06 - 19/06
- Acquired Shewanella oneidensis, Shewanella putrefaciens, Escherichia coli MFDpir for conjugation and Escherichia coli DH5a containing the pSRK-Km plasmid for Shewanella oneidensis expression
- Prepared glycerol stocks of obtained strains
25/06 - 26/06
- Purified pSRK-Km from E.coli DH5a
July
01/07 - 03/07
- Created chemically competent E.coli MFDpir for transformation
- Performed test transformations of MFDpir and TOP10 chemically competent E.coli
- Amplified BioBricks from Uppsala iGEM 2020 team
06/07 - 11/07
- Acquired Shewanella oneidensis wildtype (WT) and MtrB knockout strains, and Chromobacterium Violaceum CV026 for biological AHL assays
- Prepared glycerol stocks of obtained strains
- Amplified BioBricks from Stockholm iGEM 2019 team
- Transformed and amplified BioBricks in vivo
- Electroporation of electrocompetent cells
15/07 - 17/07
- Assembly of BioBricks was attempted, but incomplete digestion gave too high background or no successful ligations
23/07
- Restriction digest troubleshooting
27/07 - 31/07
- Acquired gBlocks of constructs or partial constructs
- PCR amplified gBlocks
- Purification of pSB1C3 for ligating gBlocks into
August
03/08 - 07/08
- Transformed to amplify gBlocks in vivo
10/08 - 14/08
- Colony PCR to select successful constructs
- Amplified gBlocks in vivo
17/08-21/08
- Started MFC with Shewanella oneidensis WT
- PCR amplified gBlocks
- Colony PCR to select successful constructs
- Transformed to amplify gBlocks in vivo
- Attempted to assemble construct from partial constructs
24/08-31/08
- Colony PCR to select successful constructs
- Chromobacterium assay to check for presence of AHL
- Ran Western blot to check for protein expression in constructs
September
01/09-05/09
- Transformed to amplify gBlocks in vivo
- Ran Western blot
- Colony PCR to select successful constructs
- Added LB media to MFC
- Replaced MFC media with M9 minimal media with lactate
07/09-13/09
- Stopped running MFC
- Measured fluorescence intensity of constructs to check leakiness of promoters
- Chromobacterium assay
- Ran Western blot
- Attempted to assemble oscillation constructs
- Acquired PFOS and 1,1-biphenyl, prepared stock solutions
15/09-17/09
- Colony PCR to select successful constructs
22/09-27/09
- Ran viability test with pollutants
- Prepared positive control (Ftsh1-Myc) for Western blot
- Ran Western blot
28/09-30/09
- Measured fluorescence intensity of constructs induced with pollutants
October
01/10-04/10
- Measured fluorescence intensity of constructs when induced with constructs producing AHL synthases and/or pollutants
- Ran Western blot
- Ran viability tests with higher pollutant concentrations
05/10-11/10
- Started MFC with Shewanella oneidensis MtrB KO
- Repeated viability test with higher pollutant concentrations
- Chromobacterium assay with constructs producing AHL synthases
12/10-17/10
- Ran Western blot
- Measured fluorescence intensity of constructs when induced with QS molecules and/or pollutants
- Chromobacterium assay for AHL
Detailed Lab Journals
Below are the detailed lab journal files. The first part ranges from the beginning until the 17th of August, and the second part picks up from there.