Luke H. Henslee's blog

Well, I think it is safe to say I'm not a blogger. Out of 10 weeks I've blogged for about half of them. Sorry, Vanessa. You can read about what I've been up to in my final report though (which I am about to send in). :)

I can't believe it is already week 7! I'm just getting into the swing of things and now I have to leave pretty soon :(

This week has been busy and exciting. I repurified two of my proteins and repeated a bunch of my experiments (results pending). Me and Garg (my mentor) also came up with a few new experiments to try out which include cross-linking experiments and yeast-two hybrid assays.

Wow, I have not written on here in a while. Sorry to all my faithful readers (Vanessa).

Things have been so busy here that I have barely had a chance to sit down! With my proteins all purified and ready to go, I have been doing about two in vitro proteolysis reactions a day! That is a lot of gel, a lot of digging around in the -80C freezer and a lot of data.

It is kind of funny that I have been working three weeks already and I haven't even purified all the protein I need to start my experiments. Who knew separating our one desired protein from the over 2,000 normally expressed in E. coli would be so time-consuming?

Our YjbH protein, which we worked so hard to express and purify, aggregated which means it is now useless. Right now I am attempting to squeeze a little bit more YjbH out of our lysate pellet and I am waiting for my gel to stain as we speak (as I type?).

Yesterday was such a blast, Vanessa is so great at coordinating fun and very informative trips for us! After testing our public transportation skills to get to the river bank, we took an awesome tour of OHSU's new campus building. The labs are nice and shiny and they are working on some really cool stuff over there.

This has been a fun and busy week for me. After starting to get more comfortable with the lab and a lot of the techniques they use here I've been working a little bit more independently.
The SDS-PAGE gel I ran after the Ni-NTA column purification turned out really well. Large amounts of Spx were detected along with a few other mysterious bands.
To get rid of these impurities I used a Hi-Q column to create a KCl gradient for ion-exchange chromatography. Using the automatic fraction collector really reduced my time in the cold room, which was nice :)

This first week has been very exciting and and informative. After being bombarded with journal articles, slide shows and lectures on various aspects of my project, I began to understand the gist of it.