Saturday, July 19, 2008

The program is ending is about two weeks. I'm sad because I definitely wish I had more time for research, but that's the way it is. I would be troubleshooting quite a few things, but the next weeks will be quite busy enough because I'll be getting most of my data then. After I get the data, I'll need to make the usual poster and Powerpoint presentation that goes along with such summer programs. I'm not nerve-wracked about presenting yet, but I might be when it comes to the actual day. First, I need those results! I recently got three different lines of flies(it took some time to create the mutant lines with the required markers, although I wasn't the one making them), which all have lacZ labelling the product of puckered (puc), which gives a measure of the Jun kinase (JNK) activity. One line is a wild-type control, and the other two are different alleles of raw, a mutant that affects larval dendrite morphology and the JNK pathway, in addition to some other affects. The raw mutants are homozygous lethal and the embryos die early, so these lines also have GFP, so I can label the heterozygotes and identify both genotypes. I need to stain these lines with certain antibodies labeling the neurons we are interested in, puc, and GFP to label the heterozygotes. I'll be spending some time trying to get good images of the patterns of expression in each line. The microscope for this takes some getting used to, and I'm not there yet. I did get to have a quick look at the control and mutant with my supervisor doing the imaging. It's pretty exciting. Raw mutants seem to have upregulated expression of puc. There is normally just one specific neuron that faintly expresses puc. The mutant is really messed up because raw also affects the development of the cuticle. The epidermis needs to close dorsally for the cuticle to exude from there, but in raw mutants it never does. It's a slow process to get these pictures because there's an ideal range of ages and orientations that I'm looking for, so it's good that I'm relatively patient. Wish me luck in finding some good embryos :)

Wednesday, July 9, 2008

These past few days, I have had a ton of fly work. I've been making crosses and collecting virgins. The nice or bad thing about fly work is that it comes in waves, but I'm very happy with how my fly work is progressing. I'm almost at the point where I'll be getting results. Skip this if you don't want the details- I'm using PiggyBac transposons to generate small deletions of specific genes. By crossing two lines of flies with the transposon inserted at different locations, and then using flp-recombinase to induce mitotic recombination, the small deletions will be generated. This takes some generations, and I need to some way to identify the progeny of the fly, from which the event happened. Some of the events are easy to identify, such as knowing that flies with white eyes have the deletion, but for others, I need to make many crosses and from those crosses, hopefully identify some for which the recombination event took place, which is the point I'm at now.

I've been disappointed with my other work, but at least it has taught me a lesson, which is to be very careful. I was generating a mutation in a plasmid with my target DNA inserted into it, and at the end I needed to digest away the original plasmid, so that I could transform cells with the mutated DNA, but I forgot to, and ended up transforming cells, which picked up the plasmid without the mutation. When I was checking to see if any cells had the mutated plasmid, I found only negatives, and that was very frustrating. At least, now I will start over and attempt to be as thorough as possible.

This 4th of July I went to Fisherman's Wharf to see the fireworks. It was pretty foggy, so some of the fireworks were colored fog, but I did see peace signs and smiley faces. I also went to Muir Woods to see the redwoods last weekend. The trees are so tall and amazing. In the gift shop, they were selling redwoods and sequoias to grow yourself. Anywhere with a coastal climate. I wish I could take one or more back to Ohio.

Tuesday, July 1, 2008

Hey everyone,

I've been working almost a month, so I have a lot to say, but hopefully, it's not too hard to read through. First, where I am for the summer. I'm in San Francisco at UCSF's Summer Research Training Program (SRTP). I'm doing research in Yuh-nung Jan's lab, working with a postdoc here. Very generally, I'm working with fruit flies, Drosophila, researching dendrite maintenance. It has been a very enlightening and fun experience. At the beginning, I did quite a bit of paper reading to get some background in the area. I have two main parts to my project and have a good variety of things to do. I'll admit that when I got my own box of vials of flies to take care of, I was excited and kind of proud, but now I have quite a few to monitor. I've also done some molecular biology like PCR and restriction digests, but I've also gotten to do some antibody staining to image neurons. It took me some time to get familiar with everything going on in lab, but my mentor has been very good about explaining the whys of each step I'm working on. I suppose this is enough about lab for now.

The program itself has some seminars in the morning. Either about a general grad school topic or a scientist comes in to present their research and how their career path went. Someone from their admissions committee came in, and I found it informative, especially since I know little about the process of applying, and I always like hearing the research presentations.

Some fun activities I've done in San Francisco so far are visiting the Golden Gate Bridge, Telegraph Hill, Fisherman's Wharf, and I went to a free Lifehouse concert in Golden Gate Park. There are also many good places to eat. San Francisco is such a fun city because there's always something to do, although the weather here is definitely colder than I thought it would be. Last summer, I was quite a bit more isolated. I liked the experience for the summer, but being in the city in a nice change. I didn't bring a computer with me, which is partially inconvenient when I need to go online, but I think also good because I can't get distracted on it as long. I use the work computer. Just a note in case you think you wouldn't survive without it because it is possible. Good bye for now :)