Team:Warsaw/Calendar-Main/16 October 2009

Creation of the cro-box testing devices Kamil

Tasks:  Ligation of the device with two variants of the cro-box Bacteria transformation I14018 brick resurrection R0010 brick multiplication 

Methods:  The ligation mix was prepared as follows (same for both variants): 20&mu;l purified cro-box plasmid 20mu;l purified reporter device 5&mu;l ligation buffer (Fermentas) 1&mu;l T4 Ligase (Fermentas) 4&mu;l H2O  A fresh bach o chemocompetent bacteria was transformed with the ligation mixtures and incubated overnight on agarose plates supplemented with ampicillin. A fresh bach o chemocompetent bacteria was transformed with the I14018 brick solution and incubated overnight on agarose plates supplemented with kanamycin.</li> Bacterial culture carrying the Roo10 brick was resuspended in 50ml of liquid LB medium suplemented with ampicillin and incubated overnight.</li> </ul>

Mutagenesis of cI Kuba

Task: Obtain a functional, termosensitive version of cI

Results: The colonies obtained from the mutagenesis of the bistable switch created three distinc populations, demonstrating the succes of mutagenesis, as well as suggesting, that more than one point mutation was obtained (later verified by sequencing; vide: http://2009.igem.org/Team:Warsaw/results#Experimental_results)