Getting Ready for the Real Experiment - Week 3

I have been practicing on building calibrations curves. This is important because the curve is used to calculate the concentration of tested samples. I have no problem with the leucoberbelin blue calibration curve which is used to calculate the concentration of KMnO4. So far the results were all very similar. On the other hand, the hydrogen peroxide (H₂O₂) calibration curve varied in every trial. A lot of noise came out while reading from the spectrophotometer with H₂O₂. My mentor thinks that the concentration of H₂O₂ is too low. On Thursday, I ran another H₂O₂ assay with higher H₂O₂ concentration and lower concentration of Amplex-red for the working reagent. The fluorescence came out with less noise. The result did not fluctuate within the concentration as much as before.

This week I made some minimal media A to be used in the assays. On Thursday my mentor and I tried running a H₂O₂ assay in minimal media. By doing so, we could measure the concentration of H2O2 in the minimal media after introducing a culture of bacteria (putida).

Besides the work I did with my mentor, I also worked with Roli Garg and Christine Romano. Christine taught me how to make an agarose gel for gel electrophoresis to separate DNA fragments. Christine expected to see DNA fragments in the 1500s. Only one of the gels came out as expected, so Christine repeated the electrophoresis with the DNA on the gel that did not showed any fragments.