Team starts re-culturing the strains in more nutritious LB media.
Prepare 50% glycerol solution for storage and autoclaved water as well
as the M9 medium. Caroline continues to work on scripts and outreach
videos. Growth of all strains is significantly improved.
Brauer Group has not had success with chromophore plasmid yet,
redesigned the plasmid by piecing together different components.
Started with cloning of proposed hybrid promoter and sent in for
sequence verification.
Had our third worldly Wednesday at Gyro House- Mediterranean cuisine.
Week
Six- July 7-13
After series of culturing testing, the best media, optimal nitrogen
source, optimal carbon source, optimal temperature for culturing are
all determined. C-source: Glucose. N-source: Glutamate. Culturing
Media: LB. Testing Media: M9. Temperature: 30 Celsius (Anaerobic).
Brauer group continued work on chromophore plasmid. Started design of
experimentation without the chromophore as a sanity check. Ptet
promoter seemed to be giving our plasmids issues as it did not
fluoresce as expected. Contacted Tabor group for possibly shipping the
plasmid to WashU.
Had a mid summer progress report meeting with the professors and
explained to them our work’s progress and difficulties we faced.
Had our fourth Worldly Wednesday at Asian Kitchen- Korean cuisine.
Week
Seven- July 14-20
Before the formal examination (Acetylene Reduction Assay) for the
nitrogenase activity, the team performed plasmid extraction and gel
running to confirm the existence of plasmid inside the strains.
Brauer Group worked on cloning for degradation tags and other side
projects. Continued conversations with shipping plasmid to WashU and
also found a promising Biobrick part and ordered from the registry.
Ben started working on a math model of our system with Brandon.
Week
Eight- July 21-27
Rebstock group ran the Acetylene Reduction Assay with GC machine for
all strains including negative control. Specific procedure can be found
in protocol.
Brauer group received the biobrick part and started cloning a new
plasmid for the chromophore part, and sent in for sequencing.
Ben continued work on math model with Brandon by sending necessary
information to him and writing ODEs.
The team had our fifth worldly Wednesday at De Palm Tree- Jamaican
cuisine… spicy!!
Week
Nine- July 28- August 3
Continued Acetylene Reduction Assays for all strains and controls.
Brauer group continued troubleshooting of chromophore plasmid. And sent
off degradation tag plasmids for sequencing as well. Designed an
experiment for light induction by co-transforming chromophore plasmids
and light regulation plasmids. Make necessary plates for experiment and
started growing cultures for experimentation.