Team:Calgary/18 August 2010


Wednesday August 18, 2010

Chris's first gel with a variety of CpxP, ibpAB, and construction of I0500-I13507
Chris's second gel with a variety of CpxP and ibpAB
Chris and Emily's gel with a mixture of digested parts including I0500, CpxP, ibpAB, and I0500-I13507


Today I spoke to Paul, Dave and Patrick about my ideas on the modeling project that I decided. We had a 3 hour debate on the topic and we came up with a different plan. We plan to make our model such that it predicts the concentration of inclusion body formation based on the free energy and temperature of the cellular microenvironment. We are still haveing issues as to how to pull of this model but we are gradually progressing towards building a concrete one.


Today, I ran several gels. The first gel that was run had CpxP, ibpAB and the constructions of I0500-I13507 and was run with a Fermentas 1 kb Ladder. The lanes were CpxP (Lane 2, 5, 6, 12, 13 on Gel 1 and Lane 4, 5, 6-10 on Gel 2), ibpAB (Lane 3, 4, 7-11, 14, and 19 in Gel 1 and Lanes 2, 3, 6, 11-19 on Gel 2) and I0500-I13507 (Lanes 15-18 on Gel 1). The gel results are shown on the right. After this, restriction digests were run of CpxP, ibpAB and I0500-I13507 along with a control of I0500. The digests were then run on a 1.0% gel which turned up empty. We suspect that the DNA was fried due to a high exposure time that was not noticed until too late. Then, we ran digests of the parts of I0500, I13507, CpxP, and ibpAB before running them on a gel. Finally, I transformed the parts of R0011 (lacI-repressible promoter/IPTG-inducible promoter), ipbAB (stress-activated promoter), pSB2K3 (for cloning purposes) and constructed I0500-I13507. Emily and I also made kanamycin-chloramphenicol and chloramphenicol plates.


Today I made kanmycin and chloramphenicol plates. I also ran a gel of digests of I0500, I13507, CpxP and ibpAB in order to verify that we have the parts that we think we have. Today I also diluted biobrick and malE BBK primers. Raida

Today, I countinued working on the Ethics paper.