User:Timothy L. Foley/Notebook/refolding matrix/2009/10/15: Difference between revisions
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== | ==Almost there== | ||
frikkin finally... it takes for ever to make these buffers... | |||
finished off the pH 6.5-9.2 buffer sets... | |||
from my back of the envelope calculations, I determined that I would need to add 0.2 mL water to each tube to get to 9.5 mL total volume...so I did this. | |||
other notes | |||
used 87% glycerol instead of 100%, so the folding reactions will be 17% glycerol instead of 20% | |||
the amino acid solutions are 1/2 the original desired concentration (550), which makes them 275 mM. | |||
We'll see what happens... | |||
==down to arraying these in 96 well plates== | |||
<BR><BR>here's what we're gonna do: | |||
array the solutions out in 800 uL whatman polypropylene (deep well) plates as storage plates for the buffer array... | |||
they come a packs of 5, so i will set up 5 of them, but only add reducing agent and ligand to one, so it doesn't get burned out before it's time to shine... | |||
the wells claim to hold 800 uL, but in reality, this volume leaves a large bubble on top (surface tension) so I figure a safe amount would be 600 uL (700 is just below the surface of the plate)... this gives, tentatively, 6 folding experiments from one stock plate... that can have reducing agents and ligands exchanged at will... ROCK! | |||
--- | |||
to do this, I need to add: | |||
534 uL each solution to each well, and then suppliment the entire plate with reducing agent (6 uL) and ligand (60 uL) before go time. | |||
get this done. | |||
--- | |||
this I can do. | |||
Revision as of 14:30, 15 October 2009
 Refolding Matrix
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Almost therefrikkin finally... it takes for ever to make these buffers... finished off the pH 6.5-9.2 buffer sets... from my back of the envelope calculations, I determined that I would need to add 0.2 mL water to each tube to get to 9.5 mL total volume...so I did this. other notes used 87% glycerol instead of 100%, so the folding reactions will be 17% glycerol instead of 20% the amino acid solutions are 1/2 the original desired concentration (550), which makes them 275 mM. We'll see what happens...
down to arraying these in 96 well plates
array the solutions out in 800 uL whatman polypropylene (deep well) plates as storage plates for the buffer array... they come a packs of 5, so i will set up 5 of them, but only add reducing agent and ligand to one, so it doesn't get burned out before it's time to shine... the wells claim to hold 800 uL, but in reality, this volume leaves a large bubble on top (surface tension) so I figure a safe amount would be 600 uL (700 is just below the surface of the plate)... this gives, tentatively, 6 folding experiments from one stock plate... that can have reducing agents and ligands exchanged at will... ROCK! --- to do this, I need to add: 534 uL each solution to each well, and then suppliment the entire plate with reducing agent (6 uL) and ligand (60 uL) before go time.
--- this I can do.
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