lysing S. cerevisiae cells

tim stearns stearns at leland.Stanford.EDU
Tue Jan 18 17:43:34 EST 1994


In article <2hhl71$r3b at apakabar.cc.columbia.edu>,
Daniel Zabetakis <dan at cubmol.bio.columbia.edu> wrote:

>4) Follow the grinding under the microscope. You should see the cells as
>ghosts or visibly broken. If the cytoplasmic debris isn't quite concentrated
>(lot's of little jiggling stuff floating around) then you need more
>concentrated cells.
>
>5) Glass tubes? Typically, I vortex 1 ml with 1 ml of beads in a 15 ml
>glass tube. Some people think plastic isn't hard enough to break cells.
>I think they are nuts, but who can say?

Checking in the microscope is definitely a good idea.  I typically get
about 75% breakage with glass beads in a glass tube.  Also, I don't
think that the problem with plastic is hardness, it's the glass beads
that are doing the grinding after all, but rather the shape.  Most
plastic tubes that people use (eppendorf tubes, 15 ml falcon tubes)
are conical, whereas most glass tubes have round bottoms.  It seems
trivial, but I think the grinding motion is better with round bottoms.

Tim Stearns
stearns at leland.stanford.edu



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