Team:Calgary/8 August 2010


Sunday August 8, 2010

Chris's gel today of the constructions of K135000-I13507 (Lanes 2-4), K239000-I13504 (Lane 5) and K239000-I13507 (Lane 6)


In the lab on a Sunday again! Made overnight cultures of the 8 colonies that showed correct sized bands from the colony PCR performed on Friday. These cultures will be plasmid prepped first thing tomorrow morning.


Late Sunday nights in the lab are the best! Tonight I finished the construction to try to get PCR [purified (linear) malE and malE31 into the psB1AC3 and psB1AK3 vectors. I transformed these into E. Coli TOP10 cells and then plated on C and K plates and left for overnight growth. I also made overnight cultures of my I0500-I13504 and I0500-I13507 constructs. These will be miniprepped tomorrow and then verification digests will be performed before we perform the Arabinose Induction trials again. I also set up two more gradient PCR reactions of malESSdel and malE31SSdel. We are hoping to get enough amplification from these that we can proceed to a PCR purification in order to try to construct these (hopefully) biobricked genes into biobrick construction plasmids. Tonight Chris and I also made overnight cultures of TOP10 cells in order to start making more TOP10 Competent cells tomorrow.


Started late today, I basically prepared the pRFP plasmid switch for the second time. But forgot to account for the fact that NEB and Invitrogen buffer numbers do not correspond together. Opps. I will try it and see if it works and in the mean time will reconstruct on Monday.


Don't we love coming to work on Sundays? Especially until very late in the evening? Today, I ran a gel of different parts that were observed in the freezer. The parts were past constructions of K135000-I13507, K239000-I13504 and K239000-I13507. The gel can be seen to the right. As well, I finished the constructions of CpxP1 and 2 with AC and AK plasmids from the digestions prepared August 6. Finally, Emily and I also made overnight cultures of Top10 Cells to start making Top10 Competent cells tomorrow.