Yesterday was entertaining, and I have more pictures for you to be underwhelmed by! Of course not, one of them is quite neat.
On arriving at the lab the membrane was retrieved from the cold room where it had rested in milk overnight, and we continued the washing with antibodies. As each wash took an hour I did some reading about yeast two-hybrid systems, which seem to be a pretty neat means of testing for the expression of two proteins which can interact with each other by affixing various gene activation and DNA binding domains to them. While I doubt I'll be using that in this lab it helped me understand a little more of these sheaths of papers I'm totally reading instead of going on facebook and playing endless spider solitaire.
On a related note, as well as reading up on FtsA I've been chatting to the post-grads and masters students who share working space with the Dafforn lab on occasion. As well as being really good eggs they're also working on some pretty interesting things, such as testing whether a protein is involved in oxidative stress responses by bleaching the hell out of bacteria with some peroxide. I'm pretty certain a post-grad course is for me.
Once the membrane was suitably covered in milk and protein, we turned the camera and controlling laptop on in the dark room. Here is a picture of the camera:
I've got to be honest, it does look a little bit like a coffee machine, only with more radiation hazard stickers. The lens is up at the top, and if you open the red door there's an adjustable tray on which you put your sample. An initially disconcerting number of switches on the front contain white light, UV, Power, and other such useful functions, whilst different lenses are available for different settings. We were using no lens and basic white light, which is good, as those UV-opaque faceguards looked uncomfortable.
We also made up a solution containing hydrogen peroxide which interacts with horseradish peroxidase to produce a signal detectable by the camera. After dumping this onto the membrane for 2.5 minutes Dr J transferred the membrane to another sheet of clingfilm and placed it into the camera's chamber, before taking several rather clear pictures which I shall show you later on. In the meantime, here's a picture of a tornado:
This is me making up a fantastic four litres of liquid broth, which will be used to grow bacteria in. I added 80g of the stuff and then the water, although really I should have added the flea first, as this is the bit that is responsible for the stirring. Several attacks with a long plastic rod later it was whirring around at the bottom merrily as an alarmingly deep whirlpool emerged. I had a while, so I took the above picture. All I could have added were two Galleons and the soundtrack to Pirates of the Carribean 3...
Embarassingly I mistook the foam at the tip of the funnel for some undissolved LB and Dr J had to turn the stirrer off to demonstrate this was not the case. I shall have to guard against being hypnotised by swirling liquids.
Once the broth was made I decanted it into 5 flasks and bunged them up, ready to be autoclaved. Nothing sterilises liquid broth quite like a good autoclaving, apparantly. I'd imagine a pressure cooker the size of a laundrette drum would sterilise most things.
Ultimately it was a pretty productive day; Dr J got a nice set of photographs showing his protein is being expressed nicely in his recombinant E. coli, and I got to make a storm in a 5-litre beaker. Not to mention the training for when I do all that myself.