continuous glycerol gradient - (Feb/26/2008 )
Hello everybody, I am actually looking to a protocol to do a continuous glycerol gradient 15-30%..Someone can help me?
you need a gradient former. the heavy solution (30%) goes in the outlet chamber (mixing is in this chamber), the light solution goes in the far chamber. pour a little heavy solution into the tube before opening the far chamber to the outlet chamber. allow to pour (slow enough that the gradient is not disrupted), with mixing, until solution is finished pouring (you used the right amounts to fill the tube to the proper level).
mdfenko says as it is; I like to add that air bubbling during pouring the gardient is to avoid; we use a long needle (for spinal liquor) to pour the gradient; to check gradient quality add phenol red in one of the stocks;
if you have no gradient mixer do a step gradient