Week 3 – Troubleshooting
(I was supposed to post this over the weekend, but the website kept lagging so here it is now!)
Since the extraction method I used last week didn’t work, I decided to try using a different plant extraction method this week. I cut my leaves into small pieces with a pair of scissors, and instead of putting them into the blender, I ground them with a mortar and pestle.
Unfortunately, when I took the filtered extract to my advisor, we were once again unable to see anything except cell debris.
I decided to look online and see if there were any other protocols for glandular trichome extraction from mint leaves that didn’t require a cell disruptor. Most protocols used cell disruptors, but I found one for RNA that used a vortexer and glass beads, which I am planning to try out next week. In addition, I was able to get a dye for staining, so it’ll help me see my target products from the enzyme extraction more clearly.
Besides troubleshooting enzyme extraction, I also worked on developing GC-MS methodology this week. Since the GC-MS is the machine I’ll be primarily using to test my results, I have to develop a protocol to use the machine that fits my project. I started off by testing for the volatiles in my mint plants, but the results were a bunch of noise (a.k.a. the results showed nothing). My advisor speculated that was because the concentration of my compounds was too low, so next week I’ll be trying this again but with a more concentrated sample.
I also tried looking for the lowest detection limit of one of my target compounds, S-limonene. This was to test how low concentration the machines (GC-MS) could detect the compound, and I was able to get some preliminary results.
Next week is going to be more troubleshooting, so hopefully it goes more smoothly than this week!