We are truly disappointed that we still couldn't make our finO and finP biobricks. We found a lot of issues that must be solved in order to achieve this objective, and we are totally running out of time =(
We decided that we are going to leave finO and finP aside for a while, and we will focus on constructing our modified version of Paris 2007 Team's biobricks.
We made a whole schedule for that, and we hope we would complete it in a week tops. The next step would be it's insertion into genomic DNA by homologous recombination.
Marcelo
YeastGuard
Dephosphorylation - CIAP test
Today we performed 5' dephosphorylation of a biobrick vector (previously digested with XbaI and SpeI) with CIAP (Protocol 10) in order to test the reaction efficiency and compare with SAP protocol (Protocol 9).
We did two ligation reactions, one using the phosphorylated vector (control) and another using the dephosphorylated (Protocol 11).
We transformed the electrocompetent E. coli (protocol 3) with the ligations and plated in LB+Amp+Kan media.