Margaret (pling) wrote,
Margaret
pling

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Concentrating!

Today I've been very busy at work. Makes a change - I've had bugger all to do for ages but today I've had too much. Can't win ;)

I've been concentrating some conditioned medium that we're going to purify a TIMP from - it doesn't express well, even in over-expressing cells, so we're starting with 15l of medium. This is a little difficult to work with - so we're firstly concentrating it down to about 2l. So far I've got 11l concentrated to 5l, so it'll take a while - tomorrow I'll work on buffer exchanging about 3l of that so I can start pumping it on the column while the rest concentrates.

I've also been doing more to our transfection. The G418 won't kill the CHO cells we're transfecting - so we can't assume that the transfected cells are infact transfected until we've killed the water controls. So we're diluting out again as we figure it's a cell concentration problem - the antibiotic has to be internalised to have an effect, so obviously with more cells there'll be fewer molecules per cell and so it'll not be as effective.

And I had a meeting with Gill about my discussion. A very positive meeting which was cool :) We went through and she pointed out bits where she didn't understand what I meant or where she thought I maybe should add something more explantory. But we ended up crossing most of those comments out coz after we'd talked aobut it she saw what I meant. Which is mostly what a viva is - discussing what you've written. So if I'm convincing then it's ok :)

And now I'm knackered.
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