08/10/2014

Today ‘s debate day to formulate our strategies for the ligation of our biobricks with professor Spartaco Astolfi.

We calculated the tconcentration of each cut biobrick and we had:

BB Essential cut with EcoRI + SpeI = 2ng/ul

BB for Bioremediation cut with EcoRI + XbaI = 1ng/ul.

BB E0840 cut with EcoRI + XbaI = 5ng/ul

BB K346004 cut with EcoRI + XbaI = 20ng/ul.

To develop a methodology is important to the ligation system to have the same amount of vector /DNA linerazed and fragmented in order to be inserted. O BB Essential (at 2ng/ul) will be ligated to BB for Bioremediation, to BB E0840 and BBK346004, in which BB Essential will become the fragment inserted in each of the linearized vectors (cut with EcoRI and XbaI).

Regarding the amount of base pairs BB Essential is two times smaller than E0840, 5 times smaller than BB Biorremediation and approximately the same size as K346004.

With these information we concluded that the amount needed of DNA for each ligation system is:

Based on the amount needed of DNA needed for each ligation system – Always trying to get an even amount of molecules –we made our very own ligation system:

We’ll digest it tomorrow