Trip Tomorrow?

This is going to be a really quick blog, but I've been so busy in the lab!  I determined today that the final step in getting the amplified DNA product ready for sequencing is too difficult to do here at Wofford with the equipment available, so I am going up to the center tomorrow to try to put my first sequencing plate in for this gene.  I've heard that it may snow tomorrow, if so I may be stuck here.  If that happens to be the case I'll just amplify more exons!

I think my thumbs are bruised.

So much PCR! Thus, so many tubes.  And those suckers hurt my thumbs when I have to open and close them so often.  But as far as actual progress goes, I started doing PCR on the exons.  I ran a gel on Exon 1 to make sure everything was ok . . . there was a lot of primer dimer so I cut the amount down again for my next batch.  We'll see how that one turns out.  I've got the 3rd and 4th exons cooking in the thermal machine right now.  Here are some pretty gel pictures for you; the bright band is primer dimer, the lighter one is the actual DNA.

My work day in the lab is from about 9am to 7 or 8pm so far.  The funny thing is I don't even mind!  I guess I just really enjoy this stuff.

Frazzled, but learning the ropes

Today was the first day I did work in the Wofford lab.  It really is a challenge to learn your way around a different lab with different equipment.  My first PCR took forever because I was trying to locate and organize all the correct pipettes and tips, etc.  It also didn't turn out well but I'm using wax to coat the next batch for testing the annealing temperature.  It's in the machine right now, I'm going to run a gel on it shortly.  I'm really, really, REALLY hoping it works because if it doesn't then serious revision must be done. I'll let you know how it goes!

This is the area in which I run my gels

This is the area in which I put together my mix to amplify DNA

Upside Down and Backwards

Everything that I did in the GGC lab today was just that.  I was a little rusty getting back to doing things; I put the PCR plates in backwards on accident and then later ran my gel upside down for about a minute before realizing it.  Everything turned out alright though!  I did my temperature gradient on the primers with a random male's DNA, then ran the PCR products on a gel.  After this, I packed up all my materials for the research and drove back to Wofford.  I don't think I've EVER been so careful driving.  So much DNA and research material . . . it's precious cargo.  Once I got back I labeled and put up everything that needed to be refrigerated/frozen.  I also wrote down an inventory of everything I brought.  To succeed with this research, I must be organized!

That's all for today, hopefully tomorrow I can get a lot done with initial PCR and gels.  I'm off to make a protocol and schedule!

I've got a fly on my face!

Ok so I titled this entry as so because when Dr. Moss was helping me set everything up today in the lab I helped his clean out some fruit fly bottles.  We were using paint brushes and at one point I accidently pushed the plastic netting back, let go, and thus catapulted a few fruit fly larvae onto my face.  This, I thought, is the life of a bio major. It made me laugh.

Anyway, I've decided I'm going to make a protocol to carry around with me in the lab so that I can reference it while I'm doing my work.  This will help avoid mistakes, I do believe, which during the scientific process can be costly and time-consuming.  And we all know how limited time and money are!  I'll check back with you tomorrow when I drive to the GGC again to pick up all my supplies and do a temperature gradient on the various primers.

Challenges already!

Today I traveled to the GGC to get the materials to start working . . . but there was much more preparation in store than just grabbing some polymerase and fleeing the scene.  Upon arrival Melanie and I immediately set up a binder for the gene I'm working on.  The binder is already fat with information, and I haven't even started working!  She explained to me why this gene was chosen for screening; afterward we set to finding all the tubes of patient DNA we're going to use.

All the tubes of DNA are kept in a cold room.  And I mean cold.  After trying to pull singular tubes out of all the different boxes off of different racks off of different shelves, we decided to just take the racks out of the cold room and pull tubes from there.  This helped us avoid freezing.  Next came the sorting and aliquoting.  It was tedious work, but it's so important to get everything sorted correctly.  The primers came in around lunch time.  My last task was to dilute and aliquot those.

Back at Wofford I went to the lab I'm working in to make sure I had key card access.  I surveyed the equipment I'm going to use and pondered how I'm going to get all my work done in the short amount of time I have. Strategy is the name of the game!  As with any research project, thinking several steps ahead is a must.  I also found out today that I have to do gels after every PCR.  This makes sense, and I like making gels.  I just have to consult the biology professors again for instructions on what I can use here at Wofford for making and running them.  I realized this is a part of real world research:  having to accommodate for a different lab in which availability and variety of equipment may vary.  This is going to be a learning experience, and I can't wait for it.

First day! Well, sort of . . .

So today the research interim group had its first (of two) meetings.  We all went around in a circle and told about our projects.  I was really surprised at how complicated and interesting the projects are going to be!  This is not to say that a Wofford student would ever attempt to glide through interim on a self-proposed project that a goldfish could easily complete, but some of the projects blew my mind a little bit.  I especially enjoyed hearing about the research projects for the humanities.  Just because I'm eternally dedicated to science doesn't mean I don't appreciate a good dose of art or literature or history.  This is a liberal arts institute after all!  It's a good situation to be in, surrounded by such a wealth of knowledge and variety of interests.

As for advances on my project, today Dr. Moss showed me around in the lab space I have been so graciously lent for interim.  I shall be returning to Room 101, where I had my genetics lab course.  I get to play with the PCR machine and the vortex and the centrifuge and micropipettes . . . see how fun biology is?  I suppose it's less playing and more utilization, considering the seriousness of the matter.  My situation is no longer that of a student trying to successfully gel electrophorese their own DNA.  Research that the Greenwood Genetic Center does legitimately helps people!  I truly appreciate the opportunities that have been given to me by both Wofford and the GGC.  I can't wait to start my work tomorrow, when I travel to Greenwood for my first weekly visit.