'Does this mean I'll glow in the dark?'

Herein marks the beginning of my final week in the lab. The eight weeks have completely flown past and I can't believe it's coming to the end-- I feel like I should only be halfway through! Even scarier a thought is that in 3 weeks time I will be starting my third year and my dissertation... Cripes.

So last time I updated, there were question-marks here, there, everywhere. There is still no definite resolution to any of those questions yet-- but work has begun to solve the mystery. Because there is a lack of Zta banding in the blots (which is our positive control band) this raises questions as to whether something in the assay causes its' degradation or whether there was something wrong with the nuclear extract I was using. The solution to this is to create radioactively labelled Zta and run it through the procedure and see if it then bands (if it does, it means the nuclear extract had degraded). If not then there is something within the assay that causes this problem and the assay can be dissected to see where the degradation occurs.

After all this, before running the procedure again, the radioactive Zta can be doped in to nuclear extract to see that the modifications have worked. Hopefully then the next individual who tackles the project can get straight to work on isolating proteins (if there are any).

Of course, it's easier said than done because firstly we have to produce the radioactively labelled [S35Meth] Zta and I'm not trained to work with radioactivity. (Admittedly the idea of working with radioactivity brings up all sorts of 'Hulk' ideas in my mind and I'm not sure I want to mutate myself yet.) The first step is to generate this radioactive Zta-- through an in vitro transcription and translation system. This requires constant testing at every stage and it quite long and laborious... But it does look like we are producing Zta mRNA which is exactly what we need at this point in time.

With this being a long process, I've been helping out with some Maxi-Preps at the same time. And it's official: bacteria smell bloody funky. I felt like I could smell it on my skin for hours later and I kept getting really worried that customers at my other job could smell it on me too (I go straight from the lab to my other job some evenings). Anyway, I've managed to maxi-prep 6 separate colonies of bacteria with different mutations-- so I think I'm adjusting to the smell a little bit.

I've also done some mini-preps and restriction digests to compare different mini-prep techniques to see which generate the best results.

So yes, the last week has been spent doing bits-and-bobs around the lab whilst waiting for the next part of my own experiment to come into fruition. This week, of course, I have to start writing my report to go alongside my 8 week experience. Slightly daunting, but obviously everyone will be able to see how that goes when they get published.