Throughout the semester I have been testing the growth rate as well a viability of Mycobacterium bovis BCG in several different media. Each growth curve requires a daily optical density measurement for two weeks. For weeks I kept on making new media varying up the recipe and calculations and finally about three weeks ago we might have stumbled upon a media that is suitable for M. bovis BCG growth in the presence of glutathione. I have conducted three growth curves using this media and plan on conducting a viability test by plating the cultures onto agar plates every day for 14 days.
After the viability test, I will perform an NADH assay, which will determine the relative NADH concentrations of my cultures. Running through each failed trial was frustrating but it taught me a lot about being meticulous and patient. Reading more literature was key to modifying the media. My next step would be to use transcriptomics to figure out which genes are up-regulated when M. bovis BCG is under non-replicative persistence. For this process, I would need to extract the RNA and then create a complementary DNA. Then I input the genetic information into a machine that scans and processes the genes. This would then tell me which genes are up-regulated.
One other aspect of my research also was to teach and help my other lab-mates with their own project. This gave me a better grasp of techniques needed to perform RNA extractions, feeding assays as well as inoculations.