Team:Calgary/7 September 2010


Tuesday September 7, 2010

Chris's gel of the PCR of CpxP and ibpAB. Lane contents can be seen below.


Today I PCR Purified malE and malE31. I then digested these as well as the psB1AK3 vector with a combination of restriction enzymes to try to get it into an AK BBK construction vector. I ligated these and transformed them into TOP10 competant cells and plated on K plates, leaving for overnight growth. I also made more K plates as well as more AK broth. Today we also all worked on the presentation that we will be giving at the 3rd annual aGEM Jamboree this coming weekend in Edmonton.


Today I ran a gel of the troubleshooting PCR. I also read the arabinose induction that was done by Emily. Although there was a mixup in samples, we still got decent readings and I will induce more samples tomorrow. I hope to get a better reading for CpxR-I13507 with MalE31 circuit.I also pitched in to work with the team for the aGEM presentation. I also reconstructed DegP-I13504/13507 with I0500-B0034 and MalE31 (Plated on AK) so that we can begin testing and characterizing the circuits.


Today, I ran another restriction digest of R0040, I13504, and I13507 to try to construct a maximum level of GFP and RFP production with the Tet-R Repressible constitutive promoter. I ran a gel to verify the PCR that was set up yesterday with CpxP and ibpAB. The lanes contain CpxP (Lane 1-3, 5, 7-8, 12-13), ibpAB (Lane 4, 6, 9-11), a positive control of Lux (Lane 14) and a negative control in Lane 15. The gel can be viewed on the right.