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Team:UC Davis/Notebook/Week 5
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Our team progressed at a pretty typical pace this week. Construction had its failures and successes as we attempted to ligate our repressors in front the RBS B0034. On the mutant side of the project we have set up the Tecan for mutant screening and will hopefully prove through fluorescence readings that we do indeed have mutants. While we are making and testing promoter mutants, we started making repressor mutants. We'll need them eventually and the mutagenic PCR's don't take too long to do while things are slow. However, testing out these mutants and finding a good range of repression is the tough part. We also cut the wild-type repressors for construction of our testing plasmid.
Finally got the Tecan set up how we want it for characterization of our mutants.
Miniprepped J61002 in a chloramphenicol backbone, this should come in handy later!
Completely reorganized the 4 degree room. Now we can actually find our plates!
Uh-oh! Our transformations from yesterday failed. Colonies on all of the controls! We're going to re-digest the following parts:
- C0040 (Cut with XbaI and PstI)
- C0012 (Cut with XbaI and PstI)
- C0051 (Cut with XbaI and PstI)
- C0051 (Cut with SpeI and PstI)
- E0240 (Cut with XbaI and PstI)
- B0034 (Cut with SpeI and PstI)
Set up PCR reactions in order to Gibson our project together. The parts PCR'd were c0012, C0051, C0040, I13458, I13453, E0240, R0010, R0051, and R0040. Will see how they look on a gel and then Gibson them together. So many primers!
Keegan took the GRE today!
Today was a really fun day! We did some minipreps in the morning but then drove down to UC Berkeley for a NorCal iGEM picnic. We got to talk with our fellow iGEMers from UCSF and Berkeley and got updates on how their projects were going. It was nice to see that there is a community of synthetic biologists in Northern California that we can share ideas and materials with. After a relaxed lunch, we had a small meeting to discuss how to achieve the medal requirements. We discussed helping each other to fulfill one of the gold medal requirements with the Berkeley team since some of our software tools might be useful for their project this year. We then got to see their lab and received some pBAD+AraC since those parts from the registry seem to be faulty. Overall a really good day!
