09/19/2014

Today we made the final test with the construction of bio sensor (BB_Essential + E0840) according to “Quantification of Green Fluorescent Protein (GFP) induced by different mercury concentrations”. The experiment was made in triplicate for each concentration. We also made measurements of those samples for its fluorescence and growth (through O.D) every hour and a half until total 4:30 hours of experiment. We had a lot of fun making the experiment! :)

We used the equipment Hidex Chameleon to measure samples with the dark plate for fluorescence and the white plate for optic density. All samples were measured in triplicate.

The results are: Optic density of DH5α exposed to 0µg/ml, 0.01µg/ml, 0.02µg/ml, 0.1µg/ml, 0.2µg/ml and 1µg/ml mercury concentrations for time 1 (1:30 hours), time 2 (3 hours) and time 3 (4:30 hours):

Intensity of Green Fluorescent Protein in DH5α induced by Hg concentrations of 0µg/ml, 0,01µg/ml, 0,02µg/ml, 0,1µg/ml, 0,2µg/ml e 1µg/ml on time 1 (1:30 hours), time 2 (3:00 hours) and time 3 (4:30 hours):

Intensity of Green Fluorescent Protein in relation to optic density (O.D.) in DH5α induced by Hg concentrations of 0µg/ml, 0,01µg/ml, 0,02µg/ml, 0,1µg/ml, 0,2µg/ml e 1µg/ml on time 1 (1:30 hours), time 2 (3:00 hours) and time 3 (4:30 hours):

Besides that, all biobricks were also digested so we can send it to the parts iGEM bank using the following system:

Electrophoretic profile of Bio bricks digestion:

1 : BB_Bioremediation in PB1C3 not digested

2: BB_Bioremediation in PB1C3 digested with EcoRI and PstI

3: BB_Essential in PB1C3 not digested.

4: BB_Essential in PB1C3 digested with EcoRI and PstI

5 : BB_Essential + BB_Bioremediation in PB1C3 not digested

6: BB_Essential + BB_Bioremediation in PB1C3 digested with EcoRI and PstI

7: BB_Essential + BB_E0840 in PB1C3 not digested

8: BB_Essential + BB_E0840 in PB1C3 digested with EcoRI and PstI.

9: BB_Essential + BB_K346004 in PB1C3 not digested.

10: BB_Essential + BB_k346004 in PB1C3 digested with EcoRI and PstI.