08/16/2014

Heeyy!! What’s up?? Today we’re going to do plasmid’s DNA extraction from the colonies that amplified the fragments of interest. Either to bioremediation, accumulation or detection.

Electrophoretic profile of the plasmid’s DNA extraction:

1 – 6: pBSK with BB_Essential + BB_Bioremediation.

7 – 13: pSB1C3 with BB_Essential + E0840.

14 – 20: pSB1C3 with BB_Essential + K346004.

Yaaaay!!! Our extraction worked! To confirm that bio bricks are actually linked, we’re going to digest to release the fragment, by doing it, we can check its size. We selected the following samples:

We used the following system:

The digestion took 1 hour at 37°C.

Electrophoretic profile of enzymatic digestion with EcoRI and PstI:

1: BB_Essential + BB_Bioremediation not digested;

2 – 4: BB_Essential + BB_Bioremediation cut with EcoRI e PstI;

5: BB_Essential + E0840 not digested;

6 – 8: BB_Essential + E0840 digested with EcoRI e PstI;

9: BB_Essential + K346004 not digested

10 – 12: BB_Essential + K346004 digested with EcoRI e PstI.

As we can see on the electrophoretic profile, the samples, BB_Essential + K346004 worked, with the PSB1C3 on the top and under it is the biobrick!

However... we have a tricky molecular question! We produced the enzymatic system to release our bio brick, but it didn’t release the BB_Essential + E0840 and BB_Essential + BB_Bioremediation (samples 2, 3, 4, 6, 7, 8)!!! Why? We thought a lot about it, and we found out! We’ll give the answer tomorrow.