Does Glass Beads Hurt Intracellular Organelles?

Namjin Chung nc1 at acpub.duke.edu
Thu Feb 13 11:41:32 EST 1997


Hi,

I'm interested in fractionating yeast cells, mainly vacuolar fraction and
nuclear fraction.  

When I break yeast cells, I generally use acid-washed glass beads
(425-600u) to disrupt cells with alternating maximum vortexing for 30s and
sitting on ice for 30s for 10-12 times.  I'm afraid if this method could
break vacuolar and/or nuclear membranes in addition to plasma membranes
and cell walls.

Can I specifically remove nuclear fractions without contamination by any
relatively easy method?  Can I pull out vacuolar fraction without losing
its content?

TIA


Namjin Chung, who knows little and asks a lot!
Program in Molecular Cancer Biology
Duke University Medical Center
Durham, North Carolina



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