08/15/2014

Our inoculum in LB liquid medium chloramphenicol 34 µg/µl with the transforming from BB_Essential + K346004 and BB_Essential + E0840 didn’t grow. ='(

We isolated from the plate with the binding made yesterday. Twelve new colonies of BB_Essential + K346004, 2 colonies of BB_Essential + E0840 and 2 colonies of BB_Essential + BB_Bioremediation.

Hoooowever, the BB_Essential + BB_Bioremediation developed!! \o/ We extracted plasmid DNA!

Electrophoretic profile of pBSK with BB_Essential + BB_Bioremediation:

1 – 2: pBSK with BB_Essential + BB_Bioremediation.

To confirm if the BB_Essential + BB_Bioremediation binding worked, we’re going to cut with EcoRI and Pstl to release the biobrick. It’s expected that the biobrick will have about to 3.057 base pairs. We made enzymatic digestion using the following system:

The samples were digested from two different clones of BB_Essential + BB_Bioremediation and we applied it in agarose gel 0,8%.

Electrophoretic profile of the enzymatic digestion using EcoRI and PstI from pBSK on BB_Essential + BB_Bioremediation:

1 – 3: pBSK “with” BB_Essential + BB_Bioremediation not digested.

2 – 4: pBSK “with” BB_Essential + BB_Bioremediation digested using EcoRI and PstI.

Definitely…. This wasn’t the expected results! =(

The enzymatic digestion should’ve released a fragment of around 3.057 base pairs, but it’s electrophoretic profile doesn’t shows a cut “vector-fragment”, but a linear vector instead. What will we do now? The strategy to solve this problin will be discussed tomorrow, because now it’s 10:30 pm and we have no way to keep working….. ZzZzZzZzZz