Team:Calgary/19 August 2010


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Thursday August 19, 2010

Chris's gel of the digests of ibpAB (Lanes 1 and 2), AK plasmid (Lane 3), and degP construct (Lanes 4-7)


Today I made a list of what figures we will need for our presentation next week. I also read over Raida's current draft of the Ethics paper. In the lab, I miniprepped last night's Overnight cultures of ibpAB-FsxA-T7. I then set up a verification digest of this piece of DNA with EcoRI and the EcoRI Buffer from NEB. I made overnight cultures of the psB2K3 vector as well as ibpAB-FsxA-T7. diluted the new malE-BBK-R* primer. Dev then helped me set up two reactions in order to amplify malE, malE31, malESSdel and malE31SSdel from plasmid DNA. We left this running overnight. Finally, I inoculated LB broth with my overnight cultures of I0500-I13504 for the Arabinose Induction assay. I finihsed making competant cells of the degP and CpxP strains from the Raivio Lab as well as the cells with our sequenced cpxR circuit. These competant cells are for testing purposes. Finally, Chris and I worked on purifying some PCR product of cpxP from a couple of weeks ago.


Today, I continued to contact companies in attempt to get sponsorships for our project. In the lab, I helped Emily make strains of TR49 and TR50 with CpxP-lacZ and degP-lacZ comptent as well as making Top10 cells containing K135000-I13507 competent. I also helped make glycerol stocks of I0500-I13504 and K135000-I13507 which were flash frozen to keep forever. I did a vacuum purification of the CpxP promoter and constructed I0500-I13507 and K239000 with I13504 and I13507. I made overnight cultures of K135000 in AK plasmid which can be used later. Finally, I did digests of ibpAB, pSB1AK3 and Alex's constructs in a gel which are on the right (ibpAB in lanes 1 and 2, pSB1AK3 in lane 3, and Alex's in the remaining lanes).


Today I worked on modeling stuff. Paul and I had a meeting about what concrete idea we want to put forth. The conclusion of the meeting was to determine and run the equations in the model by some expert biochemists and engineers before making a model of our system.