At the beginning of the first week, we gathered as a group to finalize
our summer timeline. We had to give a presentation to faculty advisors
on our summer readings.
We created team nitroGENIUS, and prepared a timeline for the summer as
well.
Later, the team traveled to Monstanto, one of out major sponsors,
Headquarters and gave a presentation to a panel of nitrogen fixation
experts on our summer project, and got a tour of the facilities.
Afterwards we started lab safety training and split into our respective
labs for the summer.
The Brauer team also started design of cloning for light regulation
mechanism and ordered primers.
Week
Two- June 9-15
Continued from Week One, the team was trained to make LB media and
completed all of the basic lab procedure training in their respective
labs. During the second week, the team also completed the LB plate
preparation. Rebstock Team members were trained for transformation of
designed plasmid to JM109, BL21(DE3), Top10, and DH5α was also done.
Started filling preliminary Safety forms.
Continued work on creating negative and positive controls for light
regulation, in addition to the light regulation plasmid and the
necessary precursor plasmid.
At the end of the week we had our first meeting with Brandon and Rajib
from Penn State to see how they can help our project from a
computational perspective; they looked at possibly implementing a
minimal nif cluster once we have the nitrogen fixation working.
Week
Three- June 16-22
Electroporation and transformation was done to all strains—plasmids
are
now in all selected strains. Rebstock Team started culturing
experimental plates and tubes of JM109, BL21(DE3), Top10, and DH5α
with
plasmids. Positive and Negative control are designed. Team also
initiated to contact Dennis Dean and two Chinese research groups about
their plasmid that can be utilized as positive control for WashU iGEM
team.
Brauer group sent positive and negative controls for sequencing and
learned how to colony PCR. Also redesigned plasmid and primers for
light regulation.
We had another meeting with Brandon and Rajib; this time giving them an
update on our wet lab work progress.
Had our first Worldly Wednesday at the Taj Mahal.
Week
Four- June 23-29
As part of outreach program, Caroline started work on the scripts of
video series introducing WashU iGEM project. Video filming is
initiated and well in progress. Rebstock Team continued culturing
the
strains growing in plates in the hot room into new tubes as preparation
for inoculation. Media used is minimal M9 media. Growth of the wild
type strains is very limited and the engineered strains achieved low
growth as well.
Brauer group continued cloning for negative control and finished
cloning of light regulation plasmid and sent in for sequence
verification. Keep troubleshooting the chromophore plasmid.
We had our second Worldly Wednesday at Queen of Sheba- Ethiopian food.
Also watched a world cup match as a group.
Brandon updated us on his progress of looking at protein domains in
different nif clusters. Considered starting a model of our system. Our
group explained to him our progress and sent him information on our
protocols
"
