Throughout the three weeks of senior project time, I have been working under the guidance of Dr. Shi Wang to move my project along. So far, we have extracted DNA from my soil samples using the Quick-DNA Fecal/Soil Microbe Miniprep Kit and assessed the quantity and quality of the samples using the NanoDrop 2000 Spectrophotometer and the Qubit 2.0 Fluorometer. These results will help us see if the samples are pure enough to be used for ddPCR. I plan to continue working at LBNL over the summer and start ddPCR tests soon, and hope to send my results to Milorganite when I am done. I also hope to expand my work beyond my project at LBNL and work as an intern on some other projects. I am very excited to see where my project and my work goes!

I have been tending to my lettuce every week since January. Interestingly, only two of my lettuce plants in the Milorganite treated pots sprouted, but those two plants flourished the most. In comparison, multiple plants (n=4-6) sprouted in each of the control pots, but their growth remained suppressed. I finished collecting soil samples on April 6th and am keeping them in a -80ºC freezer until I am able to extract DNA. I am going through the process of becoming an affiliate researcher at LBNL so that I am able to perform my extractions and the ddPCR tests in Dr. Brodie's lab next week.

Since November, I have started to collect data. I planted lettuce in 18 pots, 6 of which did not receive any treatment, 6 of which received 2 tbs of Milorganite treatment, and 6 of which received 2 tbs treatment and will receive another 1 tbs of treatment halfway through the growing process. I have collected initial soil samples and my next step will be extracting the DNA from the samples. I recently went to Dr. Brodie's lab at LBNL, where Principal Research Associate Shi Wang showed me how to properly extract DNA with some extra soil and Milorganite samples. Interestingly, the Milorganite sample did not have a high DNA extraction, which means I might have to do some troubleshooting if I want to isolate DNA from just the Milorganite again. I also have been tracking plant growth, and 6 of 6 negative control pots have 3 or more lettuce sprouts. I will have to troubleshoot a bit to see what this means, and will reach out to my mentor to see what to do about this.

In the past month, I have met with contacts at Milorganite and they have helped me establish more of the groundwork for my research. I am emailing a microbiologist and she provided me with some interesting resources about the microbial composition of their product and what the presence of certain microbes signifies about the product. Moving forward, I need to solidify some of the details of my project, like specifically what antibiotic resistance genes I am looking at and how much of the biosolid I might need. I will reach out to my mentor and other experts as well as read more research papers.

These past few weeks, I have been contacting Milorganite in an attempt to meet with their microbiologists. I am hopeful to set up a meeting with some experts that could explain their specific process of creating Milorganite. In the meantime, I am also using the resources on their website to discover more about their product on my own and think of some questions to ask. I have also reached out to my mentor, Dr. Brodie, for possible ideas. He gave me a list of companies and people to look at, and my next step will be researching all of these companies and reaching out.