Hi all! I've emphasized before how important and helpful it can be to have previous lab experience before going to graduate school. Well, recently, I've actually learned how that previous research experience can actually be a hindrance. Today I would like to post about what I'm going to call the "overly confident rotation student."

As you all know, I came into graduate school with what I consider to be a lot of research experience. I never thought of this as a disadvantage, until now. Why, do you ask? Well, let me lay some framework for why I feel this way...

All of my rotations thus far as a graduate student have been doing completely different things than I have ever done before in my previous experiences, for the most part. The first one consisted of a mouse procedure that I hadn't done before, tissue culture techniques that I'd never used, ELISA, etc.  The second one was completely different too - lots of microscopy, immunostaining for parasites, etc.  The only overlap in these were very general things like tissue culture, I have immunostained before (but never double fluorescent staining, etc.), I've done flow cytometry, and I've used a scope.  But, the actual procedures were things I'd never done.  And in both of these rotations, I've gotten As! Needless to say, Ellyn's head was getting a little inflated.

Then, a couple weeks ago, I started my third rotation, where I'm doing a cloning project and creating a deletion mutant. I was kind of excited about the cloning project! Finally, something I'm familiar with! Well, this familiarity kind of bit me in the ass.  I have been setting up my PCRs, not really paying much attention to the nitty gritty details, because I've done PCR before! I know the dilutions that need to be done for the various components. I know all about melting temperatures and elongation temperatures and times.  It clicks. I get it. I've done it. Well, I don't know about all the different PCR master mixes they use, and how they are not one and the same, and they all serve different functions. So, today I sit waiting to see if my third PCR works, because I was too much of an over-confident idiot to double check that I was using the right PCR mix in either of my previous 2 attempts. Damn it! 

So - yes, I made a stupid mistake! But, the kicker is that I made a stupid mistake twice in a row, and could have easily fixed my mistake after the first incident. But, I didn't ask questions, and that's a problem! Fortunately, the issues I am still having with the PCR don't seem to be linked to the fact that I've used the wrong mastermix.

I told the post-doc that I'm working with that I admittedly was a little too confident about my abilities with PCR, because I've done it before, and will ask more questions in the future. I personally feel that is important. If you know you did something wrong, be mature and own up to it.

So, that's my lesson for the day! Don't be the overly confident rotation student! Even if you're pretty confident about what you're doing, ask clarifying questions just to be sure. The silliest thing you can do is make mistakes that are as avoidable as using the correct master mix!

Until next time,