I try to purify a protein from yeast to assay some structural studies.
unfortunatly, the yield is really poor and I want to find some ways to
increase it. First, I change the medium and the temperature and the time of
induction but I think that now it is optimal.
Because my protein is a DNA binding protein, I think that I lost some
proteins fixed on genomic DNA that are found in the pellet during the step
of clarification just after breaking cells. Does it may be right ? Is there
a way to prevent this interactions (high salt concentration, additives ??)
I ask also, to know if there is a special manipulation to get a clear
pellet during the step of cells collecting because my pellet is rapidly
resolublize and the end of the run in spite of an optimal speed, resulting
in the loss of some cells. Is there specials additives to get clear,
compact and well packed pellet of cells, that I can put directly on the
medium (YNB -URA galactose) at the end of induction.
Nicolas pHD student Institut Curie Paris france