Week 2:
University of Iowa Department of Biochemistry

May 28, 2013

The first week of my internship at Maria Spies single molecule biochemistry lab was spent familiarizing myself with the lab and general protocols and beginning the work of creating the protein stocks that I need for my investigation of FANCJ and it’s binding and motility properties.  This first week was both very busy and very slow.  Without stocks I was unable to do much.  In order to create our protein we transfect mammalian cells with the DNA for the protein including a FLAG tag that can be used to purify it out later.  The first step in this process is to grow a culture of mammalian cells for transfection which is a week to a week and a half process.  On the other hand I had several research papers and lab papers to read and understand as well.

Everyone in the lab is very helpful with finding protocols and supplies, although I think that sometimes they must get frustrated with it since there are two summer interns in the lab and two undergraduate interns from the University of Iowa which makes for a lot of questions, but also provides a lot of extra hands for getting experiments done. When I started at the lab I was surprised to find out that both Maria Spies and my immediate mentor, Masayoshi (Masa) Honda, spoke Japanese which I started studying at the end of the school year.

The second week started out with a slight setback.  The cultures of mammalian cells that I had planned to transfect on Tuesday had not grown sufficiently for the transfection to be effective.  The transfection was carried out successfully on Wednesday though.  The cells were incubated for 24 hours after the transfection to allow the protein to fully express.  On Thursday I collected the transfected cells and froze them for storage in preparation for extracting and purifying the protein on Monday of the next week.

This week didn’t seem like it had much work in it looking at the hours spent in the lab, but most of my time was spent making sure that I understood the questions of our experiment; this is a goal that I am continually pursuing.  It is amazing how long research takes.  At the end of the second week, I had barely even started on the production of my protein stocks, let alone started my experiment.  It might be a good thing that I am planning on spending nearly 4 months here.