09/08/2014

Today we continued with total DNA extraction from wild bacteria that show resistance to high Hg concentrations using the phenol-chloroform method, started on September 6th. And guess what? We still couldn’t finish today, again! We let DNA in R buffer (Tris-HCl + EDTA) so tomorrow it will be super pretty on gel! ♥

We also continued with our new constructions and transfared DH5α with bio bricks K611025 (promoter 1), J23100 (promoter 3) e K516030 (mRFP generator) in liquid medium LB with its selection markers. (For more information, please, access September 1st and 7th)

Today we also had progress with the tests for bio bricks sequencing, started on September 2nd. We measured concentration of each part in PSB1C3 on spectrophotometer, the result was:

BB_Essential sample 10: 50ng/ul.

BB_Bioremediation sample 7: 80ng/ul

BB_Essential + E0840 sample 1: 70ng/ul.

BB_Essential + E0840 sample 2: 140ng/ul.

BB_Essential + K346004 sample 4: 160ng/ul

BB_Essential + K346004 sample 6: 240ng/ul.

BB_Essential + BB_Bioremediation sample 8: 140ng/ul.

BB_Essential + BB_Bioremediation sample 9: 150ng/ul.

We also analyzed the electrophoretic profile of each sample in agarose gel 0.8%.

Electrophoretic profile of bio bricks in PSB1C3:

1 – 3: BB_Essential + E0840 in PSB1C3

4 – 6: BB_Essential + K346004 in PSB1C3

7: BB_Bioremediation in PSB1C3

8 – 9: BB_Essential + K346004 in PSB1C3

10: BB_Essential in PSB1C3

We’re going to need 200ng/ul to 500ng/ul for sequencing. Tomorrow we’re doing the sequencing reaction!  We’re going to need 200ng/ul to 500ng/ul for sequencing. Tomorrow we’re doing the sequencing reaction! =)/p>