First week with the monkey flowers

My name is Mai Nakamura, and I'm a rising senior at East Chapel Hill High School. My PI for the program is Dr. John Willis, and I'll mostly be working with Carrie Wu and David Lowry. My project will be to figure out whether the Mimulus tilingii is more similar, in terms of its DNA and its morphology, to the geographically neighboring Mimulus guttatus or more similar to other Mimulus tilingii that are farther away. I think this method of working on two projects at the same time is wonderful for me because I not only have the opportunity to work with the molecular component of Mimulus but I also don't lose track of the "big picture" by actually doing physical gardening-type experimental work and getting soil in my fingernails which allows me to keep in touch with the underlying goals of the molecular work in the lab. I don't know about the rest of you but for me in biology lab last year, I would often know to stir this enzyme into this test tube or to pour a dye into that cocktail, but I didn't have a strong grasp of what I was supposed to be trying to accomplish. There would always be a big, gaping question - "Why am I doing this, and what is this for?". So I really like this method of two ongoing projects because it answers this question and doesn't leave me in a state of uncertainty =].

Monday: I had already met most of the people in the Willis lab last week, so Carrie and David immediately started me working on relocating various pre-planted Mimulus seeds that had been going through "winter" to the greenhouse where they will hopefully start germinating in the next week or so. With a clipboard, I wrote down the population names and the individual's number on a sheet of paper, and I later put that information on an Excel document so that the recording of the germination/flowering dates will be easier later on. Then, David and I went back down to the greenhouse and started "selfing" or self-fertilizing some Mimulus guttatus. (I found out that it's a lot harder than it looks/sounds...) Then, we went up to the lab conference room, and David and Carrie began to explain to me the concept of QTL mapping. I'm really lucky to have two mentors that are both really good at explaining hard concepts!

Tuesday: After this morning's gel electrophoresis, I went to the lab and did gel electrophoresis again with Carrie! It was great because I actually felt a lot more comfortable the second time with having this process still fresh in my mind. Hopefully, it'll be drilled into me by the time I have to do it for actual experimental data. So for the gel electrophoresis, we used some PCR results from another lab member and a hyperIV ladder...And it worked out well! We used the machine with the ultraviolet light (I don't really know its name or how it works) and printed out a picture of the gel.

Wednesday: Amazingly, some of the seeds began to germinate after only two days! However, seeds from some populations, probably those that have a long growing season that can afford to germinate later, remained dormant. The other major event of the day was "PCRing" with Carrie. It was a lot harder than I thought it would be, with all the different programs depending on the microsatellites used or whether ESTs markers are used. At the end, thankfully, all the tubes seemed as though they had about the same volume of liquid in it, so hopefully the gel that we will be making tomorrow will show good results.

Thursday: Today, Carrie and I used gel electrophoresis to test to see whether we had good PCR results from yesterday. Carrie pretty much let me make the gel by myself, always standing right next to me and pointing a few things out once in a while. While we let the gel dry, we went into the conference room again, and Carrie went over in detail what she would like for me to accomplish this summer. Since Mimulus tilingii are high-altitude plants that live all the way up in the alpines, researchers don't yet know whether they evolved independently from convergent evolution or whether there had been a widespread area characterized by cold weather way-back-when that eventually diminished to create these now isolated Mimulus tilingii populations that can only survive at high altitudes. So, what I'm going to be doing this summer is comparing, phenotypically and genotypically, the Mimulus tilingii populations among each other and between Mimulus guttatus . If we find that the Mimulus tilingii are more similar to each other, this probably means that the wide-spread colder region did exist and that there was more recent gene flow among the tilingii . On the other hand, if we find that the Mimulus tilingii populations are more similar to the various Mimulus guttatus populations, then that suggests that there has been more recent gene flow between the two separate species populations. After that, we went back to the lab for the running of the gel after which we discovered that the PCR had gone well! Once again, using that UV light machine, we printed out a picture of the gel.

Friday: Today, we did another PCR but with two different markers - EST641 and EST332. While the PCR was going, I made various dilutions of all the markers that I will be using along with some dNTP. After, I made another gel with the PCR results, and I saw that I hadn't done as well as the previous time...but it wasn't bad. That pretty much took up most of the afternoon.

I hope everyone else is enjoying themselves as much as I am! I am absolutely exhausted...