I've not updated recently, and that's mainly because a lot of the technique I've been using has been repeated over the past few days. In order to get enough protein to survive the purification etc. stages, a lot of bacteria is needed; thus I get to do lots of incubations and the such. Whilst I'm getting pretty good at it (for a second week student, anyone else here could probably wipe the floor with me), I do make mistakes. Here are some of them:
(After making a western blot)
Me: "Hmmm. This doesn't look right at all. It's...rather smeary. And I swear the band's moved down a lot...I understand as my technique isn't fantastic the line might be rather thick, but it should be in roughly the same place. Maybe the exposure was too high? No, that affects signal...argh, something's gone horribly horribly wrong. Dr J, help!"
Dr J: "Nah, it's fine. You've just got it upside down."
Then there was that time I accidentally forgot to put ampicillin into the flasks. And the time I didn't clean the virkon (bleach) out properly and incubation took SEVEN HOURS.
But ultimately these mistakes are helping me learn what not to repeat.
In other news, time to outline my project, as that's not been done yet.
Baaaaasically I'm looking at ZipA, primarily, whilst doing a little bit of work with Dr J on FtsA which is similar in that it also binds to the cell membrane and to FtsZ. The first stage is to make lots of it, the second is to purify the protein (which will result in much loss, hence the requirement for so much bacteria). After that we can do some tests on it, including perhaps crystalisation, which will be neato.
"So how's the project going?" I ask myself in some sort of bizarre self-interrogation. Well, well. The best part is seeing myself get better over time at little aspects of the techniques, whilst the worst part is the sinking feeling of dread as alarms sound and the order is given to "evacuate the facility, it's loose, it's loose". Fortunately these incidents are mostly dreams and actually everything's going as smoothly as could be expected. Next week may be a large departure from the work I've done so far and I'll be sure to write lots and lots about the fun new things I've learned how to do with machines and my brain.