July 24th, 2001

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Been busy busy at work the last coupla days. The purification stuff is winding down - I'm doing the last few concentrations and active site titrations now. Hopefully I'll not have to do this again for a while *fingers crossed*.

We're still going with the transfection too - but Nik's away, so I'm doing all the work this week. It's not too hard though, just changing medium on the 96 well plates three times this week, and keeping a couple of cell lines going in case it all goes wrong.

And I've done some more work on the project that I'm 'supposed' to be working on ... my PCR still isn't working, but Phil (PhD student with the bench next to me who's done a fair amount of PCR in the past) had a couple of suggestions. I tried one today and it didn't work, and I've done the set up for her other suggestion, so I'll see if that works tomorrow. If it doesn't work, then it's beyond my ability to troubleshoot (ditto Phil's) so I'll have to go talk to the boss. Which is kinda admitting defeat, but defeated I will be, so I might as well admit it ;)

I also read a chapter of a book that I need to incorporate into my thesis somehow ... all about exosites in MMPs, and how they affect kinetics, particularly collagenolysis. I found something to put in where Vera had indicated I needed to, and I think I don't need to mention it anywhere else - which is good, coz at first I thought it might involve major re-writes of the discussion. But I've decided I disagree with too much of what he says about the methodology so I don't particularly want to get drawn on it in the viva. Justifiably so I might add - he goes on and on about how single amino acid mutations are the only way to go. But we're talking about a 400 amino acid protein here - and all the bits are implicated in collagenolysis so where the hell do you start??? You've got to narrow it down to particular regions before you start just changing an occassional residue. But although it's justifiable, I don't want it to be a topic of conversation if it doesn't have to be ;)

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