09/12/2014

Hello!!! Today we continued with identification and isolation of wild bacteria from contaminated stream and Negro river. We made PCR of 16S rRNA with the following system:

We added 7ul on agarose gel 0,8%.

Electrophoretic profile of total DNA from wild bacteria resistant to Hg in amplified 16S rRNA.

1 – 2: Sample A1* collected from Negro River

3 – 4: Sample A2* collected from Negro River

5 – 6: Sample E1* collected from contaminated stream

7 – 8: Sample E2 collected from contaminated stream

9 – 10: Sample P1* collected from contaminated stream

11 – 12: Sample P2 collected from contaminated stream

13 – 14: Sample P3 collected from contaminated stream

15 – 16: Sample S1* collected from contaminated stream

*Resistant to 100ug/ml Hg.

Regarding to our new construction, we couldn’t extract the plasmid DNA from JM110 with mRFP generator K516030. We also transformed again to take it off the dam methylation. Could be that the bacteria isn’t efficient anymore. Because of this, we also made a pre inoculum in liquid medium LB of JM110. We used positive control with PUC72.