Phizzwizards, Ringbellers and Whoppsies!

This week I have been mostly eating Marmite biscuits…They’re a bit weird actually, but I need the energy. Unfortunately, the smell reminds me of the big bucket of yeast extract I sometimes use to make LB medium. I realise that I keep talking about smells, but the lab really is full of a variety of disgusting scents. There are shelves upon shelves of bottles each containing a glorious stench. Reminds me of the Big Friendly Giant who, I’m sure some of you remember, bottles up dreams. The lab is full of these “phizzwizards, ringbellers and whoppsies!” Just today a chemist came in and commented on how “funky” the lab smelt.

The lab has got very crowded all of a sudden! An explosion in the chemistry lab meant that all of the chemists have moved their experiments over to our lab. When I was younger, I remember thinking that science was all about bubbling flasks of colourful chemicals that would explode and fizz over, but as I grew up I began to discover that these concoctions were mainly myths. In a way, when I was told about the explosion, my childhood excitement was almost reignited. However, this was quickly snuffed out again once I discovered that the explosion was in fact just a faulty ceiling that decided to collapse…

Something rather embarrassing happened to me the other day. As some of you may know, I can be a rather jumpy individual and I was picking up some Eppendorf tubes from a 95°C water bath, when one of the lids popped open. Instead of just jumping I let out a loud shriek! Fortunately, I was the only person in the lab at the time, but I can’t believe I was scared of a popping lid! How will I be able to continue my career in lab work if I can’t even handle an Eppendorf tube?

Oh dear. My supervisor just asked me to write a research proposal to Cancer Research! The rest of the lab team seem to think it's a good idea to let me do it... They must be mad. Oh well, I'll try my best and see how it goes!



Now for a short, scientific update! I have re-done a few experiments and finally got some clear expression of my protein (see gel below)!! Consequently, I am in the middle of purifying it at the moment. This involves selecting for my protein of interest, using a flow chromatography column that binds my protein via its His-tag, therefore purifying it. Encouragingly, there was a strong peak on the graph for my elution buffer! Hopefully this means that I have got lots of my protein and not contamination, which is what I am checking at the moment by doing yet another SDS-PAGE gel (a gel that separates proteins according to their size).

SDS-PAGE

Lanes 1, 9 + 13: Soluble fractions of the 3 different plasmids I am using.
Lanes 2, 10 + 14: Insoluble fractions of the 3 different plasmids.
Lane 3: Flow through for first plasmid
Lanes 4-7: Elution for first plasmid in order of time. Lane 5 shows the strongest expression of a protein. Black line on the diagram measures it off at the protein ladder and it seems to be the expect size.
Lane 8: Protein ladder.
Lane 11: Flow through for second plasmid
Lane 12: Elution for the second plasmid in order of time.
Lane 15: Flow through for the third plasmids
Lane 16: Elution for the third plasmid in order of time.

Chromatogram