Author Archives: Mitch Lee

I’ve Only Just Begun

For the past eight weeks, I’ve had the opportunity to be a scientist. Or, at least, I’ve had the opportunity to feel like a scientist. And if I had to say just one thing about that feeling, it would be this: I want more.

I had two main objectives coming into this program. First and foremost, I wanted to confirm my desire to pursue science as a career. Secondly, I wanted to get my foot in the door so that, should I find that this is indeed what I want to do, I would be well positioned to dive in. This program, I am happy to report, has helped me to achieve both objectives.

Before this program, I could not have told you exactly why I want to pursue science. And, in fact, as I discovered during a conversation with my fellow fellow (I’ve waited eight weeks to say that), Azeb, I still can’t. But as I’ve thought about that question, I’ve realized that asking it is like asking why I like the taste of lasagna. Why do I like lasagna? Because I do.

Now, I can tell you why I want to pursue science: it both satisfies me intellectually and fulfills my desire to contribute. Let me elaborate.

A spark of excitement flared in my mind when I remembered each morning that I was on my way to lab. I found myself looking forward to the endless procedural tasks that waited for my at my bench even though I knew full-well that I was going to mess up at least a couple times while trying to tackle them. It didn’t seem to matter if I was doing my third gel-extraction of the day (which I admit was my least favorite procedure) or gathering flow cytometry data on the gene expression levels produced by a new set of plasmids that I made the day before (which I thought was the most exciting step), the spark never left. Of course, it wasn’t always sunshine and buttercups. There is nothing more frustrating than waiting around until 9:00 pm for a digest only to find that it failed utterly. But even in that moment, after a few brief moments of melodramatic woe-is-me, I felt reinvigorated by the prospect of retrying it the next day. Why? Because I do.

Now, as I hinted earlier, there’s more to my wanting to continue to research than just my liking it.  While working in the lab, evaluating data with my mentor, attending lab meetings, and reading as much literature as humanly possible during the fifteen-minute intervals between procedural steps, I felt myself part of the scientific world, a participant in the ongoing exchange, accumulation, and distribution of constantly new information. For me, though a neophyte I am, this was fulfilling and rewarding.

In my eyes, that is the about as best a confirmation as I can hope for.

Thanks to this program (at least for now): mission success!

A Brief Walk in Their Shoes…Oh, and Polyphenic Butterflies!

Having the opportunity to attend private seminars given by some of Duke’s most premiere researchers has been by far one of the most rewarding aspects of being an HHR Fellow. And it’s rewarding for two reasons: because I get to learn about really cool science (duh!) and because I get to learn about various life experiences and paths that lead these people, who at one time were young undergraduates like me, both enthralled and somewhat terrified by the prospect of becoming a member of the global scientific community, to become the investigative inquirers that they are today.

In contrast to what I expected given the often cookie-cutter profiles that my generation of prospective researchers is often encouraged to build  in order to maximize our chances of attending the “best” graduate schools, the background and developmental stories of these researchers were as diverse and their research. Some began their college careers wanting to do things as unrelated to science as something can possibly be. Others set off wanting quite the opposite. But in no two cases were the paths the same. In fact, the only constant that I noticed within each narrative was a certain element of serendipity. Now, I don’t mean to devalue the effort and struggle put forth by each presenter. I instead point out this seemingly ubiquitous fortune because its presence, and the fact that all of our presenters capitalized on the serendipitous moments presented to them, indicated to me what I think is one of the most poignant tidbits of wisdom that can be drawn from the life-story portions of our seminars: be open to and prepared for the favorable opportunities that life presents.

To me, this is a comforting message. I find it so because I am becoming more and more acutely aware of the difficult struggle that lies before me on my path to researcher-hood. I will have to work and struggle to succeed as our various presenters have trailblaze my path. It is the struggle inherent in trailblazing that frightens me most, so it’s comforting to know (or at least to think and hope) that fortune might smile down on me somewhere along the way and nudge me toward a fulfilling objective.

Ok, enough with the deep, philosophical garble. On to some cool science!

One of my favorite seminars was given by Dr. Frederick Nijhout, who studies, among many things, polyphenic development in insects. To provide a touch of background, polyphenism refers to the generation of two or more phenotypes from one genotype based on environmental conditions and influences. Dr. Nijhout’s research involves investigating both how the phenotypic outcome of polyphenic traits is determined in butterflies and how, once determined, that particular phenotypic outcome is achieved in a system designed to create more than one outcome. So far, his research has led him to investigate ecdysone and juvenile hormone, both of which seem to play an important role in mediating the impact of environmental factors on genotypic expression and in bringing about different phenotypes that result from these environmental factors.

Because my project involves the manipulation of gene circuits via environmental cues, which has sparked my interest in epigenetics, Dr. Nijhout’s research was especially intriguing.  Dr. Nijhout’s ability to explain, at least in terms of hormones, why members of the same butterfly species are either black or brown depending on the season in which they are born is fantastic. In doing so, he has contributed a small but important bundle of information and ideas to the age-old discussion of nature vs nurture. I think that’s pretty cool.

One step forward, 3 glances over my shoulder, and 1/2 a step back

Science is difficult. I said that I expected this coming into the program, and indeed I have been proven to have anticipated correctly. Oddly enough, however, I came into this program expecting the theory behind the science to be the most difficult part, not only in terms of understanding (in my case, for example) mechanistically how genes can interact with one another in a way that produces specific regulatory pattern and the implications of those regulatory patterns, but also in terms of understanding how the data that we hope to obtain will shed light on these questions.

As it turns out, this expectation was wrong. Understanding the theory behind my research has proven far more manageable than what has proven to be a far more tricky, and sometimes cruel, adversary: project design.

At first, my project seemed easy enough to plan. I knew where I was starting, and I could see and understand plainly enough the general steps that I needed to take in order to reach the goal.

Step 1: Take this promoter and put it into this plasmid.

Step 2: Take this gene and insert it next to the promoter in the same plasmid.

Step 3: Repeat steps 1 and 2 with different combinations of promoters and plasmids. What I did

Seemed like a straightforward enough task to complete to me. But little did I know. It turns out that completing step 1 alone is much more easily said than done. Not only are there a barrage of procedures required to make the plasmid (each of which introduces innumerable opportunities for error) but there are also a barrage of procedures required to make sure that the end product is the one I intended. After all, I have to make sure that I end with plasmid A instead of G, X, V, L, or any other possible mistake.

Having to plan and carry these steps out carefully enough to be confident that I am actually producing the plasmids I need has challenged me more than any other aspect of my project. I can’t count the number of times at each step of the process that my mentor has asked me “But what about this?” or “Did you account for this?” Often, the answer is “uhhhh…no” and it’s back to the drawing board. As I’ve undergone this cycle many, many times over these six weeks, it’s often felt like I’m taking one step forward, glancing over my shoulder at least three times to make sure that I haven’t messed up, and, realizing I have, taking half a step back. That’s a definite woe.

But don’t misinterpret. Making it through that process has been one of the most rewarding parts of this program. Being able to say “Ok, this is where I am now and I need to get to this point” and then being able to make an outline of procedures, one that efficiently  accomplishes the task while making the proper and needed security checks, has been an incredibly rewarding and empowering. I’ve yet to master the art (in fact I just found out that my plasmids may not function properly given that my original template plasmid seems to have a faulty promoter), but I’ve enjoyed learning. Despite the frustration of having to redo several steps along the way, I still find myself excited by the idea of doing it right.

Mantis Shrimp Attack with the Force of a 0.22 Caliber Bullet

Hearing about the projects and research being done by the other HH fellows was, in one word, exciting! And it was exciting for two reasons. The scientific theory behind each project–ranging from studying the genetic variation that drives speech-pattern development in song birds to the role of signaling pathways in the development of an autoimmune disease that causes dry eye–is, alone, enthralling and engaging on its own. However, far more striking was seeing the variation in approaches that many of the projects take. While I listened, absorbed, and attempted to comprehend the hard work that each of my colleagues is doing, I became distinctly aware of the variety of avenues the one can take through research (there is more than just endless pipetting out there!…not that I dislike pipetting, as I’ve already discussed). In this way, I guess, seeing their work as a reaffirmation of the beauty of science. The presentations I saw were a reminder that, regardless of the implications of the projects or their applications, science is more than worth doing. It’s just too cool to pass on.

Here’s an example of why.

Dani’s project, in particular, stands out to me. Dani is currently studying how cells differentiate in the appendages of mantis shrimp during the developmental phases of their growth. To begin her presentation, she explained to us that there are two basic types of mantis shrimp: bashers and spearers. Bashers have bulky claws that they use to smash open shelled prey with astonishing force (the acceleration is comparable to that of a 0.22 caliber bullet leaving a gun). Spearers instead use their smaller, barbed claws to spear passing soft-bodied prey like fish with an acceleration that, while less impressive than that of smashers, is still formidable. She further explained that while the manner in which they use their claws is different, both groups share a common physiology, having their claws on arms that are divided into four segments. Her research, however, focuses on spearers. This is because the specific development of each arm segment impacts the effectiveness of their appendage more-so than for bashers.

So how, roughly, is she studying the pattern of tissue differentiation during the growth and development of these appendages?

To begin her project, Dani took a trip with her mentor up to the Smithsonian’s Animal Archive in Virginia. While there, Dani and her mentor took approximately one thousand photos of 60 specimens (I take my hat off to you, Dani). Once she returned to her lab, she used computer programs to map key physiological points on the clawed appendages of the shrimp, points that she will cross reference between the sixty specimens to determine which segments of the appendage develop from the same root tissue and which do not. Her reasoning, as I understood it, was that if two or more segments show a consistent size ratio across a significant number of specimens, then it can be inferred that those segments of the shrimp’s arm developed from the same root tissue, or were at least regulated during their development by the same mechanisms.

Right off the bat, when Dani said that she is working with Mantis Shrimp, I knew I would especially enjoy her chalk talk. Being an avid scuba diver (and having spent the majority of my youth wanting to be a marine biologist) I am a sucker for scientific looks at ocean-dwelling creatures. After a small googling spree incited by Dani’s presentation, I have found myself especially fascinated by mantis shrimp. Not only are they beautiful, but they also have incredible physiologies (their eyes are also a marvel of nature). As such, I am excited to hear about Dani’s progress as these last few weeks of our program pass.

Minipreps, Digests, and Gels

Describing a “typical” day in the Buchler lab is difficult, as my agenda for any given day depends on the tasks required to advance my research project to its next step. That being said, there is a general pattern to what I do, although this pattern does not apply to a “typical” day, but rather to a typical “stage” of my project, the completion of which may be spread over a few or several days, depending on what the stage of the project. As such, in order to give as descriptive and revealing a look into my work as possible, I will thus devise a hypothetical agenda (comprised of tasks that I may not all actually complete on a given day) that illustrates what each of these stages may involve.

Most days begin with a meeting at 9 am for all of the Howard Hughes fellows where we either discuss important aspects of being a researcher or enjoy a presentation by a member of Duke’s faculty on the research that she or he is performing. Once these meetings are over, and usually after a quick stop at Twinnie’s Cafe for some coffee or a bagel, I arrive at lab around 10 am. Upon arriving in lab, I set up my little section of a desk with my laptop, important papers, and my lab notebook, and then walk over to my secondary mentor’s desk to discuss with him what we need to accomplish during that day. Once we have a tentative schedule of tasks laid out, it’s off to the races. Almost inevitably, this means beginning with a mini-prep (which is scientific jargon for saying that I begin the day by extracting the plasmid DNA of bacteria that have been growing overnight). This step is almost always the first thing on the agenda, regardless of what type of “stage” my research is in. This is because I almost always have to grow a fresh batch of the plasmid that I am working with for the procedures of the day.

Once I have my plasmids, my next step is almost always to run an analytical digest and analysis (I mix a portion of my plasmids with enzymes that cut the plasmids into segments that I can evaluate by length after running them through a gel that separates the segments by length) to see if the plasmids that I have just obtained are the plasmids that I want. Although–in order to keep this post relatively short and (hopefully) interesting–I will not discuss the specifics of each procedure, I will give a brief description of the specifics of this step in order to give readers who are unfamiliar with research procedures a small, descriptive sample of a typical procedure for synthetic biology.

Performing a digest and subsequent analysis involves three basic steps:

1. Mixing solution from several different vials through a lot of pipetting and letting the mixture sit in warm water for about an hour.

2. Mixing agarose with hot water and pouring the mixture into casts to make a gellatin-like substance that can be used to separate segments of DNA by length

3. Putting the gel (with digested DNA inside) into a machine that runs an electrical current through the gel for about 20 minutes (which, simply put, is what drives the DNA to be separated by length within the gel.)

In other words, much of the time I spend in lab is used pipetting, mixing, pouring, placing, and waiting. In fact, almost every procedure is a sequence of mixing solutions from several different vials, placing the new mixture into a machine that serves a specific function (be it analytical or procedural), waiting for the machine to work, and then either analyzing the output or using the output in the next sequence of pipetting, mixing, etc.

After completing my daily miniprep and digest, my agenda may look something like this:

3. Isolate a segment of DNA that I want to insert into a plasmid (I do this through a miniprep and a digest as described above)

4. Prepare the plasmid into which I want to insert the segment from step 3 so that it is ready to accept that segment.

5. Insert the segment from step 3 into the prepared plasmid from step 4 via a ligation reaction (a reaction where enzymes attach the insert to the plasmid so that it becomes one whole unit)

6. Isolate the new plasmid that I created in step 5

7. Insert the plasmid that I isolated in step 6 into a strain of bacteria

8. Grow the bacteria from step 7 overnight so that I have a fresh supply of my new plasmid to work with the next day

9. Repeat this process all over again (staring from a miniprep) with a new segment of DNA to insert

Each of these steps is part of what I call a “stage” of my research (a stage might be, or instance, taking plasmid A and making it into plasmid B) and involves procedures very similar to that for running a digest (i.e. pipetting solution together, mixing, etc.) All in all, there are probably 8 stages to my project, meaning I have to start from plasmid A and get to plasmid I by going through plasmids B, C, D, etc. If I’m lucky, I can compete maybe two stages in a week, altho0ugh that is an ambitious estimate given that I didn’t include all of the analytical steps that I have to do to make sure my synthetic steps are going as planned.

Now I know what you’re thinking…”that sounds so boring.” And, indeed, performing a miniprep for the twentieth time is not what I would call the most exhilarating experience. But I enjoy each step none-the-less, regardless of how many times I have done it. I find it stimulating to consider the theory behind the procedure itself (i.e. considering how the reagents used in a mini-prep actually serve to isolate the desired DNA). But what’s even more exciting and rewarding is to consider that every mini-prep I do, every digest and gel I run, every ligation, every step that does not lead directly to data takes me one step closer to getting results that I can see and analyze and apply. In this light, I find even the most repetitive tasks to be fulfilling. They are an essential and inescapable part of being a researcher and I spend 10:00-5:00 each day completing as many of them as I can, all so that I can be that much closer to completing my goal.

Designing bistable switches and oscillators for gene regulation in E. coli

Researchers in Dr. Buchler’s lab are working to better understand the regulatory mechanisms that occur naturally within the gene networks of organisms. The bulk of their research goes toward answering three basic questions. Firstly, how do these mechanisms function? Secondly, how do these mechanisms influence the large-scale workings of an organism’s gene networks(s)? Thirdly, how can these mechanisms be utilized or manipulated in order to give greater control over inter-gene regulation to researchers?

To investigate these questions, members of Dr. Buchler’s rely heavily on synthetic biology, which in their case involves building synthetic gene circuits–small clusters of genes that influence one another’s expression through a regulatory mechanism of interest in either a natural or controllable way–that are then inserted into budding yeast. Once inserted into yeast, these gene circuits and the effects of their substituent regulatory components can be monitored using well-established techniques and equipment common to yeast molecular biology.

Like the majority of the research occurring in Dr. Buchler’s lab, my project is to further develop our understanding and ability to utilize two powerful classes of regulatory mechanism: bi-stable switches and oscillators. But what exactly are these?

A regulatory mechanism is called a bi-stable switch if it can be used to switch “on” or “off” the expression of a gene (or genes) in a manner that is both highly controllable and highly stable (I use quotation marks because the expression of a gene can never be fully on or fully off, but rather in a state of relatively high or low expression, high being “on” and low being “off”.) In other words, a bi-stable switch allows a researcher to “turn on” the expression of a gene and have that gene stay “on” indefinitely, and then “turn off” the expression of that gene and have it stay “off” indefinitely.

An oscillator, on the other hand, is a regulatory mechanism that, once initiated, turns “on” and “off” the expression of a gene at regular intervals of time without the need for interference by an outside force (like a researcher). A synthetic biologist might, for instance, design an oscillator that turns a gene “on” for ten minutes, then turns it “off” for ten minutes, then turns it back “on” for ten minutes, and then back “off” again for ten minutes… and so on (I chose the interval of ten minutes arbitrarily).

Returning back to describing my project, I am working to develop a gene circuit that functions as a bistable switch and another that functions as an oscillator in E. coli bacteria. Although the Buchler lab does not typically work with E. coli, the circuits that I am helping to design are based on circuits that my secondary mentor, Sargis Karapetyan, successfully created for yeast. Controlling those circuits in yeast, however, proved to be more difficult than expected, so Sargis and Dr. Buchler decided to try and make use of them in bacteria, which have been shown to better cooperate with regulatory mechanisms like bistable switches and oscillators.

I cannot say exactly how we have designed these circuits, but, if our experiments and designs fare well, I’m hopeful that my work will contribute a small fraction to the writing of a research paper that reveals all.

“2 a.m. lab work was the best” -some insight from Dr. Nicolas Buchler

Preparing to interview my p.i., Dr. Nicolas Buchler, was admittedly a daunting task. After all, I had asked him to squeeze thirty minutes out of his tremendously busy schedule so that I could pick his brain with questions that were completely unrelated to the grant proposal that he was doubtlessly in the middle of feverishly writing. What should I ask in my limited time? What information or insight would be most valuable? Luckily, as our conversation transitioned from discussing the factual aspects of his career to conversing about his subjective experiences as a prominent researcher, the more important topics and questions made themselves self-evident. By the end of the interview, I walked away with three especially helpful insights that I want to share in this blog. But before I share them let me summarize (as best I can) his career and the path Dr. Buchler took to become a primary investigator in the Duke Institute for Genome Science and Policy.

Dr. Buchler–a little to my surprise–majored in physics at UC San Diego (which he sheepishly admitted he chose to attend partly because of an obsession with the LA-based band, Jane’s Addiction) . When I asked him why he chose physics, he replied that he entered UC San Diego knowing that he wanted to study science, and that his decision on physics stemmed from several factors. First and foremost, he liked the extra attention and student-teacher interactions that he had as a physics major due to physics being a much less popular than biology or chemistry. Secondly, he greatly disliked the pure memorization required of chemistry majors, especially during their studies of organic chemistry. He did, however, say that he views a molecular biology course as being a defining course of his undergrad career (which I found both serendipitous and appropriate given that Dr. Buchler was one of the teachers of the molecular bio course I took last semester). His enjoyment of this course motivated him to pursue graduate studies in biophysics at the University of Michigan, where he earned his Ph.D. while using computer simulations to model and study how proteins fold.

Up to this point in his story, Dr. Buchler’s path to researcher-hood followed a route I had more or less expected. But given the a-typical histories of several of Duke’s top researchers, I knew Dr. Buchler probably had a few unexpected steps up his sleeve. And he didn’t disappoint. After completing his doctorate, Dr. Buchler “crashed” (as he put it) a six-month genomics workshop in Santa Barbara where he met his first post-doctoral advisor. This workshop, he said, set the trajectory of his career toward becoming a genomics scientist. Having decided on that field, Dr. Buchler then decided to take a summer course at Cold Spring Harbor Laboratory that is dedicated entirely to teaching the theory, procedural techniques, and experimental designs needed for studying genomics in yeast. This class, which enabled him to dive into the world of yeast genomics, led him (thankfully both for me and science) to eventually become the primary investigator in the Duke Institute for Genome Science and Policy under whom I am now working.

Now that I’ve recounted his journey a bit, I’ll share some of the especially useful and helpful bits of conversation that I had with Dr. Buchler. Three pieces of insight/advice stand out in particular. The first is his response to my question: “what do you think is an important or essential step for undergraduates hoping for careers in research?”. His response, not surprisingly, was “do research.” He said that the increased opportunities for undergraduates to do research are a resource of tremendous importance because they give undergraduates not only the chance to confirm that they enjoy research, but also the chance to figure out which type of research they most enjoy and what field most appeals to them. He also said that doing research as an undergraduate teaches hands-on skill and how to think like a researcher, both of which will be essential tools in grad school.

The second tidbit of insight Dr. Buchler shared pertained to the transition into grad school. “Don’t expect to be a great grad student just because you were are a good undergraduate student. Some of the best grad students weren’t that great of undergraduate students and that’s because the mode of thinking in grad school is much different from the mode of thinking in undergraduate.” He elaborated that he feels this is because one has to switch from “learning mode” to “generating mode” upon entering grad school, which is (returning to his first insight) why he feels that undergraduate research is so essential. It helps students to realize whether or not they are compatible with grad school.

The last piece of insight that I would like to share came from Dr. Buchler’s response to two questions that i asked at the end of our interview. The first was, “What is the transition from being a grad student or post-doc working under another person to being a primary investigator in charge of others like? I was especially eager to ask this question because I have found myself wondering more and more if I would enjoy being a primary investigator, most of whom seem to spend the majority of their time writing grant proposals and performing administrative tasks and very little time being directly involved in the research they oversee. Dr. Buchler’s response was a mixture of comforting and disenchanting. Firstly, he said that no one really enjoys grant proposals (I cringed a little inside), but writing them is an essential component of any p.i’s job, which you eventually learn to accept. He also said that the severity and enjoyability of the transition depends largely on the person making it. Some people, having spent six years doing hands-on work, welcome the chance to solely work on theory and project design while only overseeing the hands-on work. Others keep their labs smaller so that they have time to perform some of the research directly themselves. His last reassurance was that if I enjoyed the science and work that leads up to becoming a primary investigator, that I would find overseeing and getting funding for the continuation of that work by others to be worth the transition.

Photos

photo 1   Prepping an E. coli culture

photo 3   Working on some plasmid designs (

photo (1)

More Plasmids? Please!

When we gathered for our first official morning meeting as Howard Hughes Fellows, we were given—in addition to coffee and an immensely appreciated breakfast—a few handfuls of useful advice, encouragement, and assurance about scientific research, its methods, purpose, and benefits. We were especially encouraged to consider what we expect and want to gain from our experiences as members of the program. Although this question seemed difficult to answer as I contemplated it while walking to my lab that first day, this past week has served to reveal what I desire from the coming seven weeks.

I expect a challenge.

Now, before I elaborate, I should elucidate what this past week has entailed. I am working in Dr. Nicolas Buchler’s lab under the direct guidance of Sargis Karapetyan on the development of regulatory gene switches and oscillators in bacteria. Needless as it is to say, I knew little to nothing about what the end of the above sentence meant when I first walked into the lab. The past week, therefore has consisted of a nearly endless cycle of reading, studying, questions, and explanations from Sargis (not to mention my careful yet often unproductive examination of several plasmid maps). At first the going was tough and I struggled to keep up with the theories and scientific jargon common to synthetic biology. But I slowly picked up speed until I had a basic working understanding of our objective, the methods needed to reach it, and the results that we hope to obtain. In other words, I succeeded this past week. And I felt accomplishment along the way. So, to say that I expect a challenge doesn’t quite paint the full picture.

I expect a challenge that I can overcome.

Let me expand on this. Scientific research is a brain-straining, time-intensive, and laborious pursuit that requires dedication and a deep familiarity with the subject under investigation. As such, I expect to feel a little in-over-my-head at times. Not in a desperate, “this is impossible” way, but as a pleasant reaction to the vast amounts of knowledge, skills, and experience that wait for me in the coming weeks. As I have said, building a working level of knowledge on the particular subject of regulatory genes in bacterial plasmids, let alone the rest of molecular and microbiology, has already proven difficult. I’ve read several papers of which I understood approximately 25% on first reading. Now, however, I understand them almost completely. I’ve also become competent with plasmid mapping programs that, at first, made my head spin (one very specific goal I have for the summer is to become a master of SnapGene and SeqBuilder). Because of these small, but important steps forward, I also expect that I will build a working understanding of the knowledge necessary to pursue my project. But more generally, I expect to learn about the scientific process—from idea development and project planning to execution, analysis, and presentation of the results. I expect these things not because I believe the specific outcomes of this project will be indispensable to my future, but because mastering both theory and method would mean that this program has helped me start thinking and learning like a scientist. In light of this, my general expectation can be extended once more.

I expect a plasmid-filled challenge that I can overcome and benefit from.

Finally, I expect to better understand what I will call “the people side of science” by the end of my eight weeks in Dr. Buchler’s lab. Science is by necessity a collaborative effort, so being able to interact effectively with peers (who are tremendous sources of knowledge and experience) and to contribute meaningfully to the group’s ideas and knowledge is an essential skill for a scientist. I, for instance, would sill be stumbling around in the dark were it not for Sargis, who not only supplies me continuously with the background knowledge that I need but has also helped to steer my project in a meaningful direction. Without his guidance and experience, many flaws and overlooked complications (not to mention an important pair of poorly-selected primers) would have gone unnoticed. More than anything, I have enjoyed learning from him, picking his brain, and reveling in his intellectual fervor (he, for instance, has completely changed for the better my opinion of scotch tape as a tool with a short anecdote about its centrality in the creation of graphene). By the time I leave in late July, I hope to have built a strong relationship with him, Dr. Buchler, and the rest of the members of the Buchler lab. In other words…

I expect a plasmid-filled challenge that, with the help of others, I can overcome and benefit from.