Sunday, December 13, 2009

reading, writing, and everything in between

Be skeptical.

That's what my advisor told me when I brought in a few papers a while back. And also what he said when it got to explaining my rationale behind my experiment predictions.

Part of my work involves an enzyme called AMPK, so I've been doing a bit of reading on it. But since it's such an ubiquitous protein, there's been a lot of work done on it already, even if I look into specifically what its role is in food intake control (another part of my project). So to organize my thoughts and see what parts I was still missing out on reading about, my advisor asked me to write a minireview (~5 pages) about AMPK and its role in feeding control in the brain, so that's what I've been doing in my "spare time" for the last two weeks.

Writing it was a little harder than anticipated. The first day of writing, it took me about three hours to write a paragraph. Not because I didn't know how to word things; it was because I was averaging one citation a sentence. Which means for every sentence I write, I have to read an entire paper written by someone else. I suppose I could read a whole bunch of abstracts and string them together to make an instant review, but that's out of the question. First off, my advisor is going to know whether or not I really read the papers; and second, sometimes the abstracts will sugarcoat things, and unless the reader looks into the methods, it'll seem like everything is legitimate.
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I'm also in the process of scribbling ideas to write for my written prelim exam. In general, my project has to be on leptin in the frog, but since it wasn't cloned until 2006, there's still a lot of basic info we don't know yet, like if it signals through particular pathways, or if it is produced in a certain way. These basic questions have already been answered in mammals, and it seems to me like a lot of the mammalian studies nowdays are focusing more on different brain regions that leptin acts on, how they "talk" to each other, where other signals might come from, etc. It would be pretty boring (not to mention very uncreative) of me to base my written exam off of the statement "This stuff about leptin is true in mammals. Please give me money so I can do the exact same thing in frogs."

So now I'm kinda stuck on how to use the frog as a model to advance the field. Which probably translates to "I should read a lot more..."

Friday, October 16, 2009

the hardest part

We had a lab meeting earlier this week, and I was the presenter, so I spent a good amount of time putting slides together last week. If the old rule of "a minute a slide" applies, my presentation should have been a little less than 20 minutes long.

It was an hour and a half.

I got interrupted at practically every slide.

And I didn't know the answers to a lot of the questions.

So to fill in some of the gaps, I spent a good chunk of time this past week adding, deleting, and rearranging some stuff so it addressed the comments made in the meeting, and I ran it by my advisor today. The verdict: read some more.

The slides I got the most interruptions at were the slides that described questions I wanted to address via experiments. I kept getting called on for clarifying what the question really was, what my hypothesis was, and the reason why I formed that hypothesis. It turns out that the vast majority of my background information came from studies done in mice, and while it's generally assumed that the results would be the same for frogs, that's obviously not a good enough reason to base a series of experiments off of. There were a lot of preliminary questions that needed to be addressed first, using the FROG as a model before moving on, and I didn't realize that, since I had automatically assumed that an important molecule would be evolutionarily conserved between frogs and mammals.

That being said, there's a surprising amount of information we DON'T know (or I don't know) about frogs. This information had been well-documented in mice, but I had taken this info from mice, extended it to frogs, and proceeded to the next step, without bothering to test to see if the information also held true for frogs.

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Last term in that grant-writing/prelim-giving/paper-reading class, we had a professor come in and talk to us about what makes a good grant. Three basic parts:
1. whats the question?
2. how do you test it?
3. so what? (why should we give you money?)

He said that the reason why a lot of grants are tossed out is because they don't have a good question in mind, and that the "what's the question?" part of the grants is the most difficult to answer. Originally I had disagreed; I thought "so what" was the hardest, since not everyone is going to be utterly fascinated in learning about social behavior in purple carnivorous snails (just an example). However, after the past two weeks of working on these slides, looking over at the comments made, and going back to revise things, "so what" is actually a pretty easy question to answer... once I figure out the question I want to test in my experiments.

So I'll be writing up a list of things that need to be answered, and try to weed out which question is the most fundamental...which will be the starting point for subsequent months.

Monday, September 7, 2009

faith in science

I was laboring on Labor Day; apparently my cloning experiment didn't work again. The concept is simple enough: cut plasmid/inserted gene, purify, and then glue together. So why has it taken me over two months to do this? (aside from losing the DNA, breaking machines, fighting the Incredible Shrinking Insert...)

Molecular biology (or science in general) at its best (read: textbook figures) is a great concept. We have machines we didn't have a decade ago, protocols that have been refined multiple times, and reagents that can be mass-produced so that they aren't nearly as expensive (although $150 for a tiny bottle of Taq is still way too expensive, for my taste), but even with the technology we have now, everything is far from foolproof. PCRs will just fail for no apparent reason, and the only "logical" explanation was that whatever day the experiment was done on was just a "bad lab day." Likewise, the first sentence in our lab protocol for ligations (where two pieces of DNA are glued together) is "sometimes it works, sometimes it doesn't," making it seem like you should mix the necessary ingredients together, follow the guidelines, and *hope* that it works.

Science is like a religion in itself; sometimes things just happen and nobody knows why. I've been restriction digesting and purifying "my favorite gene," which is supposed to be 500 basepairs long. Somehow after purification, the gene "shrinks" to 300 basepairs. I've ruled out every possiblity I can think of, and there's nothing that can really be done except start over (and maybe whine about it for a while). Yes, cloning sucks, but I suppose I just have to *believe* that whatever I'm doing is the right thing, and that one day after tapping my foot 55 times while wearing a red tshirt and singing along to some given song on my iPod, I'll get my ONE much-needed clone.

Seriously though, howcome superstition isn't more blatant in the research field?

Saturday, August 1, 2009

Information Overload

It's generally a good idea to read up on papers related to your research topic as preparation for prelims, and a rule of thumb is to read about two papers a week. My advisor told me about two weeks ago that I could probably be more ambitious than that and read a paper a day.

He has a good point, since my undergrad career consisted of a wide smattering of classes with no particular emphasis on one subfield in bio. But a paper a day? I'm not sure I can absorb all that info at once; I have to read a paper at least twice to get the most out of it. The first time I read it, I skip the methods section and all the figures. Second time around I'll actually look at the pictures, and (maybe) come up with more questions.

Hopefully I get better at info absorption over the next month; once school starts, my schedule is gonna be insane.

Tuesday, July 7, 2009

Year 1 in retrospect

I've been meaning to write this post for about two weeks now... first off, I finally decided to stay in this current lab (working with frogs). That doesn't mean I'm not intimidated by my PI; I've gotten a bit used to it, and plus, being a little more intimidated by this PI than the other one would push me to work a little harder and really know what I'm doing.
That being said, I don't think I'll ever get over this "being terrified of my boss" phase. Hopefully by prelim time? If not, it better be by defense time...
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Several weeks ago, one of the professors down the hall was talking about the learning curve in grad school. He said it's pretty amazing how much of a difference one year of grad school makes on a student's reasoning, presenting, and writing skills. It's hard to say where the learning curve is the steepest, but I would guess somewhere in the first two years. I was archiving my rotation presentations and written summaries the other day and before putting them away, I was reading through them. Frankly, my first written summary in grad school sounds like an 8th grader wrote it. :-/
It's gotten easier for me to see the connections between theory and experiment design (something that took me literally forever to do in my undergrad lab), but I still have a ton of stuff to learn. I thought of a possible thesis project about three weeks ago, and talked to my PI about it. There doesn't seem to be too many people working on that particular topic as far as my PubMed searches tell me, but I suppose I could go off on a tangent and search for somewhat related topics...

Tuesday, June 16, 2009

doing science properly

Last term I took a grant-writing/prelim-giving/paper-reading class, and one of the sessions was dedicated to writing a good grant. The guest speaker was a professor in the department who has never had to resubmit an NIH grant in his 30+ years of tenure as professor. He brought in his experiences as a grad student at MIT and how there were two professors there who did science very differently, but both ways were ideal, depending on your personal style.

The first way is to go off on crazy tangents from one root project. It's like saying, "mutant A has an obvious eye defect as heterozygotes, but homozygous mutants for A die." You could have someone in your lab working on what this eye defect is and how it relates to the mutated A, and another person working on the timing of death. Why do the homozygotes die? What stage? What went wrong?

The second way is to keep working on one central question, discovering things that relate to it along the way, but never fully answering the question. Going back to mutant A, you could ask "why do homozygotes die?" Then you could find out that they have defects in gene A, which is part of pathway B. But that doesn't mean pathway B is the ONLY one in the system that is making mutant A unhealthy. In other words, is there anything working with pathway B? Does it act alone? Is there a mutant A that doesn't have problems in pathway B, but has problems with receptor C instead?

The unfortunate part about getting grants is that the government expects neatly-laid out experiments, with not much wiggle room for failure (one of the main laments of one of the professors I talked to). But since NIH grants are stretched over a five year period, how is anyone supposed to have the insight to know that some project that looks promising now won't be a dead end? How do you account for pop-ups that are related to your project, but are completely unanticipated?

Science is already a competition for funds, but the restrictions on what you can write in your proposal and what you can't possibly foresee makes it more and more difficult. We can't possibly outsource research, but will the sheer difficulty of getting money drive down the population of scientists?

Friday, June 5, 2009

panic

Recently I got an email from the PI of the zebrafish lab I worked in last summer, saying she heard from the grant review panel, and now she has sufficient funds to support another grad student. I'm also finishing up the rotation project in this current (frog) lab, so now the question is where do I want to go for the next 4+ years?

Both PIs are tenured professors in developmental biology, working with different model organisms. They give good mentoring advice, are both stationed on the third floor of NatSci, and are well-known in their fields. When it comes to the important parts of grad school (like what kind of guidance you're gonna get), they're pretty much identical.

So now I think the question becomes "who am I a little less intimidated by?"

Here goes...

Thursday, May 14, 2009

on being old and boring, part 2

The fear of being boring seems pretty common; I was talking to one of my friends who is in grad school for chemistry in Japan, and we were discussing the difficulty of meeting people outside our given departments. Not just for fun, but for the sake of diversity in conversation topics.

Since lab is such a large component of the doctoral program, it's impossible not to think about it even when I'm not there. And since a large part of research seems to be troubleshooting, conversations with other students center around what could be done to fix things, or getting advice with dealing with advisor expectations, etc. I think for the past two weeks that I've been out of classes, most of my in-lab conversations circle around "where do we keep the pipette tips?" or "my Western Blot flunked. What now?"

Last term I took a seminar-based class called "Teaching in the Sciences," and as a so-called "midterm project" we were supposed to put together a powerpoint presentation on something not science-related. (gasp)

The instant the professor gave us that stipulation, we all panicked a little, as if saying "but I don't know anything outside of science! I don't even remember what hobbies I have besides from PCRing and pipetting!"

I guess my non-lab related work is cut out for me this summer.

Sunday, May 10, 2009

on being old and boring

Before I entered grad school, I'd hear all these horror stories about life in the lab. For example, one lab I heard about had an air mattress in the electrophoresis room, in case people were there late at night running gels. Another lab had a student who eventually got addicted to NPR because there was nobody in lab while he was working. I'm not addicted to NPR just yet, since this current lab doesn't listen to it, but I think I'm becoming addicted to my iPod.

My schedule for the past week has been pretty much the following:
10am to 5 or 6pm: lab
5 or 6pm: walk home and eat dinner
after dinner: either go running, then analyze data, or read up on what I'm supposed to be doing the next day
etc..

Unfortunately the majority of the time I spend in lab is spent in silence (save for the music from the earphones). One of the grad students is prepping for prelims, so he's probably not going to be in lab until those are over. The other grad student is in and out, and its pretty sporadic. There's a few undergrads, but they come in pretty sporadically as well.

Hopefully it gets a little better as the summer progresses.

Back when I interviewing with various schools, I was telling my friend Dan that I was afraid of either losing my sanity (gradually) or regressing back to shy (or something among those lines). He laughed at me and said he couldn't imagine that happening.

Seriously though... spending 7 days a week doing labwork means it's on my mind 90% of the time, which would severely limit the number of conversation topics. Grad school is making me boring, and I need to find a new hobby.

Or go on some sort of roadtrip.

Sunday, April 12, 2009

what recession?

People say that Ann Arbor (or UM in particular) is like a giant bubble: whatever is happening outside usually doesn't have too big of an effect on what's inside. We didn't need any financial bailout; the unemployment rate in the Deuce is 7.5%, one of the lowest in Michigan. Professors enjoy job security, because we can't possibly outsource scientific research or our education.

I guess this would be more true if I were still in undergrad.

The lack of money for research is really taking a toll on many of the PIs here; with thesis mentor decision deadlines looming, everyone is scrambling to find a permanent home, but who has the space to take on more students? I've done four rotations, and I enjoyed three of them, wanting to go back and do a thesis, but the labs either have grants pending and won't know until later in the year (when it might be too late to join) or can't afford to take students.

Kinda makes me wonder if this "economic stimulus" is just a myth, since I don't see any of it happening anytime soon.

Grad students are either paid as research assistants or teaching assistants. In MCDB, the teaching requirement is 2 terms, and I think during those two terms, the grad student's stipend doesn't come out of the professor's pocket. So an extra teaching load is another way to join a lab that might not have sufficient funds, but I talked to the MCDB advisor, and she said it was not recommended due to the time spent away from the lab bench, and it'll drag out the time until graduation.

So.... onto rotation #5. There's so much more to learn... just like the first one this past July. The project is definitely interesting: stress hormones and its role in development (how ironic, given my own cortisol levels right now), using Xenopus as a model system.

Never worked with amphibians before... better start reading up on metamorphosis.

Wednesday, February 25, 2009

Fourth and final

Started my last rotation today.

I was doing pigmentation and adaptation (in particular, starvation) in fruit flies during the previous rotation for January and most of this month, but this new lab is dealing with neuron development. Still in fruit flies, but a whole lot of microscopy and larvae dissecting. Not to mention cell bio... I really need to brush up on cell signaling.

I've had no motivation to do work lately.. maybe it's because it's the middle of "spring break" (applies to undergrads only, unfortunately) or maybe I just want to settle down in a thesis lab and get started. I have a feeling that choosing a thesis lab is gonna be difficult: out of my three past rotations, I'd like to go back to two of them. So right now it's cell fate determination in zebrafish vs. evo-devo in flies. Tenured professor vs. relatively new professor. If this rotation goes well, it'll end up being tenured professor vs. relatively new professor vs. REALLY new professor.

On choosing labs, I'm getting mixed messages. My dad's telling me to go with a tenured professor; someone's who's established in the field. They'll have lots of publications in their name and they'll have an "easier" time getting funds, he tells me. On the flip side, younger PIs have a stronger drive to publish, since they're still establishing themselves in the field and/or are subjected to the tenure review process. Either way though... I know that all the labs I've rotated in so far have good mentors.

In a different note, I didn't realize how many people in grad school are older and more experienced than me; it seems like not too many people came straight from undergrad. Many have had at least one year off from school and have done lab tech work for quite a while. Others are in their upper twenties/lower to mid-30s, are married, or have kids already. It kinda makes me wonder if applying and entering grad school was a hasty decision on my part.

Then again, three years from now I guess it won't make too much of a difference, since everyone's going to have candidacy (hopefully) and working on their projects.

Rotations are exhausting. Must decide by Tax Day...