Monday, 19 July 2010

When the going gets tough, the tough steal an obselete cooling device from another room and shove it between the wall and a disrupter

Pretty, isn't it? I think it goes well with the tubing.
This is an FC-360 cooler thingy, and as its instructions were not on the supplier's website Dr J and I had to guess, based on a later model, which buttons would be less likely to result in a smelly inferno. It worked surprisingly well, cooling the lysed cells down to 0.5 degrees celcius. Oh, yes...the cells...

Look at them! All frozen and such. To thaw them, we suspended them in breaking buffer, a cocktail of tris and other such lovely chemicals. Then we stirred them for some time until it held the consistency and appearance of mushroom soup (deceptive yet inedible lab hazard). Then protease inhibitors were added to prevent the cells from digesting themselves once lysed. Unfortunately these turned the goo into what resembled crab soup in colour and smell (deceptive yet inedible lab hazard). It stunk most of the lab out rather quickly. Nasty stuff.

Anyways, the disrupter was pretty effective, turning the mush of defrosted bacteria into a thick soup of cellular contents and fragmented membranes. As it was only the membranes we're interested in, they were spun at 10,000rpm for 30 minutes, and then 35,000rpm for an hour. After removing the supernatant, what remained looked a little like what you get if you leave a red fruit pastille in your mouth until it's a bare slither betwixt your tongue and your palate. Unfortunately it still reeked of crab soup (deceptive yet inedible lab hazard).

Whilst the membranes were being pelleted in a snazzy new machine with a delightfully retro interface (think the old school thumb-scrolled wheels on ancient white-numbers-on-black-squares alarm clocks), Dr J showed me how to set up a load of yeast wells for culturing. Tomorrow, I'll be taking the optical density, doing some crazy (simple) maths to work out how much to transfer into the new set of wells, and brewing up a set for western blot analysis. I'm sure it'll be a lot of fun, and hopefully I'll not grow anything lethal by accident. Although a Zombie Apocalypse would make for a great report, I doubt the novelty would last.

I'd give it about 28 minutes.

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