08/16/2014

Today, because of our previous results, we decided to do a binding test with our T4 DNA Ligase. We linked the K346004 cut with EcoRI and Xbal, to itself, to analyze the possible alternatives of linking it to pSB1C3.

Electrophoretic profile of the binding test of K346004 to itself.

1. K346004 cut/linear with EcoRI and XbaI not linked.

2. K346004 cut/linear with EcoRI and XbaI linked.

As we can see on the electrophoretic profile, T4 DNA ligase is active! Knowing that, we concluded that our binding system made on August 12^th worked, but the DNA concentration was too low to grow transformed bacteria that would show in their plasmids the desired linked fragments.

Due to this result, today we concentrated the binding system of BB_Essential + BB_Bioremediation, BB_Essential + K346004 and BB_Essential + E0840 and we transformed DH5α by electroporation.