Week 5

Well, we're coming down to the wire here in terms of finishing up my experiments for my presentation at the Symposium as this, the penultimate week, wraps up.

The first few days, all I did was prep and run an experiment. Simple enough, as I've done it countless times before. I (in some cases we) split cells, treated cells, collected lysates, poured gels, loaded gels, ran gels, performed a western blot, treated the membrane, so on and so forth. This experiment finally left me with some usable data in terms of what I can put in my presentation.

Thursday, things got hectic. The data which was collected Wednesday needed to be followed up on by another set of experiments. This, along with the stages of two experiments that will be run in the next few days. This had several experiments occuring all on one day. I was on my feet almost constanly, finishing collecting lysates here, exposing film there, taking pictures of cells there... Certainly an eventful day.

Even though I'm staying late today, and possibly Monday and Tuesday, it's really going to be a rush for me to finish my presentation and experiments by Wednesday morning, but with luck (and Dr. Kridel's help) I should be able to manage it.

Week 4

The 4th week was a short one, only 3 days in-lab due to the 4th of July. Though I enjoyed the day off, this did leave me with 4 days of work to do in an actual timespan of 3 days.

Tuesday was incredibly busy. I did three Protein Assays, loaded and ran two gels, performed two western blots and collected a set of Lysates. That means I was running two experiments simultaneously. This was the only day I had to do this, because the results of my third Protein Assay showed that there were problems somewhere in the experiment (possibly in the preliminary stuff that I had no part in) that spoiled both our controls, and rendered the experiment unusable.

Wednesday and Thursday were uneventful. I treated the membranes from Tuesday once each day (peIF2-alpha on wednesday, and total eIF2-alpha on thursday), but this left me about two hours of work to do, and the workday is much longer. So during the down-time I helped our grad-student, Joy, with several of her projects. This involved collecting Lysates (a set of 8 and a set of 12) and doing some tissue-culture. This also got me a crash course in ER (Endoplasmic Reticulum) Stress and how it applies to cancer.

So the fourth week was short and uneventful, but well spent.

Week 3

Week 3... Halfway done...

More of the same this week, with the exception that my work actually related to the presentationI'll be giving at the Symposium. The big thing this week was doing a pair of experiments from front to back (meaning all the way from Tissue Culture to Membrane Treatment) at the same time. Though it actually ended up being more like 1 1/2 experiments.

The first experiment went well, and the data was what we were expecting (maybe "hoping" is a better word...) and our second treatment designed to check for accuracy of the other treatment came back with excellent results. We even got to messing with the standard procedures a bit due to how well it was going. The second one, however, taught me the pains of research... the hard way.

The Tissue Culture, in which we treated several plates of LNCaP (Prostate Cancer) cells went well, and we let them incubate for several days. The lysing and collecting the lysates went well. The gel setting, loading and electrophoresis went by without incident. Then came the western blot. I loaded the clamp backwards, meaning that I put the gel on the wrong side of the membrane. Such a simple, stupid mistake messed up and entire day's work.

And then, it gets even better: I tried the experiment again the next day, and got through it without incident, except for one little problem. We didn't have enough sample to actually do the entire experiment. So our results were muddy, unclear and didn't help at all. Well, at least I won't be making that mistake again, and as Dr. Kridel says, "worse things have happened in the world." I just have to do the experiment again next week.

All in all a pretty good week, despite that small problem at the end.