09/04/2014

Growth of bacteria with our bio bricks in medium LM (LB + mercury chloride) it’s in progress!!! DH5α transformation with bio bricks didn’t work! We made the transformation again, DH5α with BBa_K516030 and BBa_I712074. We’re hoping that no it will grow!!!

In parallel we have the tests with mercury and the new constructions. We’re also isolating wild bacteria from the stream contaminated with industrial residuals and from Negro River. Yesterday we transferred to solid medium LB and the result was:

Wild Bacteria resistant to high Hg concentrations from Negro River – A1*, A2*

Wild bacteria resistant to high Hg concentration from stream 40 (contaminated with industrial residuals) – E1*, E2

Wild bacteria resistant to high Hg concentrations from stream 40 (contaminated with industrial residuals) – P1*, P2, P3

Wild bacteria resistant to high Hg concentrations from stream 40 (contaminated with industrial residuals) – S1*

*Resistant to 100ug/ml

Today we made a test for kanamycin (kanamycin 50ug/ml, tetracycline 12ug/ml, ampicillin, 100ug/ml, streptomycin 20ug/ml and chloramphenicol 34ug/ml) of those unknown bacteria (samples A1, A2; E1, E2; P1, P2, P3; S1).

Today we also had progress with our tests for bio bricks sequencing, started on September 2^nd (For more information, access September 2^nd). We made a new PCR with different concentration of PSB1C3 with BB_Essential + K346004. We used the primers VF2 and VR.

PCR test from PSB1C3 with BB_Essential + K346004

*Resistant to 100ug/ml

1 – 6: Sample 10 from PSB1C3 with BB_Essential + K346004 on respective concentrations and/or volume: (1) 0.5µL non diluted, (2) 1µL non diluted, (3) diluted 1:5, (4) diluted 1:10, (5) diluted 1:25 and (6) diluted 1:50

7 – 12: Sample 11 from PSB1C3 with BB_Essential + K346004 on respective concentrations and/or volume: (7) 0.5µL non diluted, (8) 1µL non diluted, (9) diluted 1:5, (10) diluted 1:10, (11) diluted 1:25 and (12) diluted 1:50

Now we could amplify! =) We’ll keep doing the quantification and tests to obtain 200ng/ul – 500ng/ul, the necessary concentration for sequencing.