Week 9

8-5-09
-Inoculated all mutant strains and WT into 2.5mL LB broth @10:10am @RT
-Finish final touches on Furse paper
-Continue Final paper
-Continue final presentation work

8-06-09
Gathered samples from a nearby river, two of them field-tested LBB positive.
Inoculated strains were mixed into different concentrations of Fe (1,2,4,8,16,32 mM) and then left overnight. We are checking for differences in siderophore production, but C1-32 oxidized significant amounts of manganese and the precipitates were found on the side of the tube. F1-32 also came up LBB positive, but the C1-32 liquid did not. This means essentially all of the oxidized manganese was left on the side of the tube after ~20hrs, which indicates that all of the manganese was oxidized. There appeared to be too much manganese for simple contamination alone to account for it all. We are repeating the procedure with strains C1, F1, and WT at all concentrations of Fe so as to check if in fact the level of Mn in Trace Elemental Mixture and what is normally in the cell was in fact all precipitated.

8-07-09
The Fe assay is not going well, basically the mutants seem to randomely be able to oxidize manganese, and our current theory is that there is some contamination in the test tubes we are using. Future experiments should acid wash all test tubes before assuming there is no manganese left in it. The seemingly random oxidation patterns suggest contamination, which due to human error would also appear essentially random. This perhaps marks the end of my bench work, although I believe we are going to try one more experiment on Monday and then officially call it.