Issue Sixteen, this thirty first day of July, 2003

'A simple scream of rage is sufficient. You scream and you leap'

Back in the first issue I reported that Sauron said something about using PCR primers to delete an internal sequence, and the sequence you want to delete is the primer, or something. I think I know what he meant, and he's talking about overlapping multi-primer extensions.

Which just goes to show, it takes nearly half a year to understand anything he says.

I actually had to get up and leave last week. Sitting at coffee, and he's ribbeting again about how we do stuff. You know, 'We should think about how we do things.' Well, we have and we do. And we've pretty much got it sorted, you know. 'These protocols were written when blahblahblah'. Yes, and now when we do a protein prep we don't spend all day doing it, do we? And we can do other things at the same time, can't we? And our protein doesn't piss out all over the floor because the equipment leaks, does it? So we haven't written timings into the protocols because (a) we're flexible and (b) we're not anally fucking retentive.

Then he talks about saving time (hah) and that maybe (watch out, here it comes) ammonium sulphate precipitation after ion exchange would be a good way of concentrating the sample and of course you're doing gel filtration next so you desalt 'for free'.

I made my excuses and left. I mean, furrfu, it's two thousand and bloody three and he's still pushing a technique that admittedly is quite useful when you're starting with two pounds of mashed up chicken gizzards but today, for recombinant proteins, there is a Better Way and I'm really getting quite pissed off. I did try AS once for a protein prep about three years ago and it was bloody useless; he doesn't understand that just because we don't do what he says doesn't necessarily mean we haven't thought about it and tried it and found it to be shit. Likewise, our old favourite beta octyl wotsit was waved at me this week. I'm setting up some more screens and he wanders over and suggests, almost plaintively, that I try it. I tell him I have and that everything fell out of solution. So he says to use only a little. 'I waved the bottle over them,' made him go away again.

Haven't heard 'convenient restriction sites' for a while, but I was shocked to find, whilst looking something up, that I had used that very phrase in my thesis (1994). But closer examination revealed that I used it in the opposite sense to Sauron's way: I said that PCR primers can be used to introduce convenient restriction sites. Phew.

Maybe it's because I'm going on holiday tomorrow and hence my patience is just about exhausted, but there don't half seem to be a lot of stupid people around these days. I made a protein calculator from Perl and put an interface to it up on the interweb (email me if you want to see it), and made it available to the entire department. I'm generous like that. Anyways, I get a couple of emails from people saying, 'wow, this is great, can I cite it in my thesis?', blah blah blah (modesty prevents me from relating any more), and then I get a complaint.

Apparently someone thinks that my calculations are wrong, because they're getting an answer a hundred fold more than the BioRad machine is giving. I puzzle over this for an hour, ask to see his input, write a few lines of code to log what people are putting in, and generally bang my head against my Mac in frustration. I go and make a cup of tea (yes, I'm doing this in my own flaming time, at home) and I suddenly realize what the problem is.

The problem is that this person is a complete berk. He actually comes to see me the next day after I email him saying what his problem is, and even then I have to explain in three different ways that the number he is getting from my program is correct, it's just adjusting for dilution into the cuvette so that the answer he sees is the concentration of his original sample, not the concentration in the actual cuvette.

'But that's not what the BioRad machine does'.

I DON'T GIVE A FLYING FUCK WHAT THE STINKING MACHINE DOES, I'VE GIVEN YOU THE RIGHT ANSWER YOU THUNDERING MORON!
I didn't say that. But I thought it. And then I had to explain, in words of one syllable or fewer, exactly how one goes about calculating an extinction coefficient and then how you get the protein concentration from that.

Huh? I mean, don't they teach this stuff in schools? I bet even my three-year old could figure it out.

And then of course, I've left in my lines of code logging what people enter, and I find that some twonk is pasting in nucleic acid sequence. Maybe it's not obvious that it's only for protein, thinks I.

Nope. I couldn't make it any more obvious. So now the code checks if the user has input only A, T, G, C, or N, and returns an appropriately rude message. It's a learning experience all right. I keep learning that people are more stupid than I give them credit for.

I need my holiday.

Good news, Sauron flew to the US yesterday, I go away Friday, he's back Monday; and when I get back two weeks later he's off for a month. Woo hoo! I'm going to pay for it though - we're scheduled to fly to Florida end September. Obviously Sauron thinks he alone can't scare our collaborator enough, and needs me there. I am _dreading_ the flight. Does anyone know where I can get a week's supply of morphine? (For me, not him). Hmm . . . maybe there's a reason we have a Medic in the lab . . .

More good news, the mRNA I glommed from the Red Headed One gave superb RT-PCR results. Mwah, thank you thank you thank you.

Mental Emails

Strange email from someone at University of Washingtion, saying they've got mail for me from someone in Minnesota. Very strange. Lots more mental email this week, but I can't really be arsed. I'm going on holiday.

Lab Rats

Grasshopper _still_ owes me a beer for that reference.

I was going to rant about sodding Studmuffin and his sodding MP3s, but I can't really be arsed. I'm going on holiday.

Enjoy the summer. BBQ when I get back?

Going on holiday,

Richard (- did I say I'm going on holiday?)