Week 3:
University of Iowa Department of Biochemistry

June 10, 2013

This week was referred to as “transgenic week” by the lab members due to the fact that we injected sperm nuclei (which had been modified by the reaction of the nuclei with two of our linearized plasmid constructs) into eggs this week.  And when I say eggs, I mean about five hundred for each construct.  Tuesday was injection day and instead of labeling lots of tiny micro-centrifuge or PCR tubes and pipetting small amounts of liquid between these small tubes, the day was spent using tiny, glass needles back-filled with the REMI reaction mixture and a syringe pump, to poke relatively big holes into about a thousand eggs.  After that experience, we then had two sorting periods where we took the dead embryos or unfertilized eggs out of the Petri dish so the death signals from those sick cells wouldn’t kill the few that had successfully been fertilized.

I’m proud to say that my first injection period (although I took about 3 hours compared to the 3 years of experience guy’s 1 hour) I was able to get about 70% of Joes surviving embryos in the first day.  Yuan, Sarah, and even Dr. Baker were impressed that I was able to attain a number fairly comparable to the graduate student’s success rate.  The members of this lab have been unbelievably supportive and understanding and have made this experience overwhelmingly positive.

I also started working on construct H34 by creating the H34 fragment via PCR.  I came in on Sunday to sort tadpoles and begin the digestion of H34-fragment 1 (H34-f1) in preparation for ligation on Monday. Yes, you read that right.  Tadpoles.

After injecting the REMI reaction on Tuesday, not even a week later they already resemble tadpoles and swim around when you stimulate them with a pipette.  It blows my mind that they develop this quickly and that I got to see it happen from the moment we fertilized them.  Now we’ll get to see next week if the gene we inserted is being expressed in at least 10 of them.  The transgenic process practiced in this lab isn’t very efficient, but it’s fast and relatively cheap so we just inject about 500 eggs and hope that 10 of them express our gene.

The FUTURE program offered some meetings this week for the undergraduates.  One meeting went over the graduate school programs offered by the University of Iowa which gave information on who to talk to at what point, what to include on the applications and when to ask who about how to apply, and the various advantages the U of I graduate programs had over other graduate programs.  The presentation was given by Dr. Spitz, a member of several admissions boards and oxidative stress in cancer biology and toxicology researcher who was very vibrant and able to show his excitement for the programs and his research.  There was also an informal reception allowing the undergrads to mingle and talk to one another where I met Brianna from our very own Cornell for the first time as well as some other students from Luther.  We had a very good time talking about the projects we are working on this summer and also having a few laughs about non-research-related things as well.

This next week we’ll be injecting two more constructs while I continue to work on H34.  There is a lot of work to do, but it’ll be no problem with the continued support and guidance from the lab members.