Starting out, I worked on a potential side project involving the honey bee. It is a bit of a pet project of Josh's and definitely has my curiosity piqued. I will "bee" looking at genetics in the honey bee populations around the valley, and determining a sort of family tree for them. How closely related are honey bees from varied locations around the valley? This also leads very well into studying the hybridization (africanization) of the local bee population, which is what has my interest.
I constructed traps last semester from 251mL (20 oz) soda bottles. I cut the top off of the bottle at the point where the curve of the neck meets the barrel of the bottle and inverted it (without the cap) in the bottle. I then poured in a layer (not reaching the former top of the bottle) of sucrose solution in one, and humming bird nectar in the other and placed them in the PC garden in close proximity to flowering plants.
The traps in location
While waiting to see how well the traps worked, Josh and I began to work out the DNA extraction protocol from a new kit from Zyma Research, their DNA mini-prep kit. We attempted an extraction with a couple samples of crickets (rather than wait for the bees). The protocol was relatively straight forward, and we did get DNA. Unfortunately, this wasn't the most fruitful extraction. It appears that the DNA had been sheared. I believe that this is a result of one of the first steps. The samples were placed in Zyma's Bashing Bead tube and then vortexed to agitate it. This essentially lyses the sample. The protocol stated this was to be done for 10 minutes and did not specify the intensity of the agitation. When I run the next samples, I will do one at a low intensity for the full 10 minutes, and another at high intensity for a shorter period of time. This should give me some insight as to the best way to do this or if this step is even what resulted in the shearing.
Approximately one week after setting my bee traps, I found three bees. All three were in the trap containing the hummingbird nectar. This makes me wonder if the sucrose might be more effective with the addition of food coloring, and also which colors may be more effective. So, these bees will be my first subjects and I will be using them for my next round of DNA extraction.
During this same time period I also did a test run through of a different DNA extraction technique for my continuing Palo Verde research. I mentioned in previous posts the kit that I had been using was yielding less than satisfactory results and that I may scrap it for another method. I did so. I am now using an extraction kit from Sigma-Aldrich. It is a far less complicated extraction method and saves me a whole lot of grinding with mortar and pestle. It is more of a "fast and dirty" technique but should suit well for my purposes. The first extraction did yield DNA, but not real well. It was no fault of their system it was complete operator error. I will be doing the extraction again tomorrow.
My DNA extraction setup
Here's to hoping for another successful semester!
Is it wrong that I am already apprehensive about presenting at the S-STEM conference at the Estrella Mountain campus at the end of the semester...
Is it wrong that I am already apprehensive about presenting at the S-STEM conference at the Estrella Mountain campus at the end of the semester...
