Preparation of nuclear extract: How to lyse the cells?

Julie Gervais 3000JGER at VM1.ULAVAL.CA
Wed Mar 22 21:38:19 EST 1995

Hi everybody!

  I'm having a hard time trying to reproduce the protocole of Wang et al (in
Biochemistry 1992- vol.31 p.3694-3702).  My cells are just not breaking open!
In this protocole, they say that, after a digestion with zymolase (1mg/g of
cells), the spheroplast preparation "was lysed in a motor-driven homogenizer
with 10 strokes "(!!!).  They do not talk about the device itself and, so, I
tried to break them using a polytron (maximum power, more than 5min...) from
Brinkmann cie ( model Kinematica):  it just do not work!

Do somebody know what type of apparatus they used?  May I ask for tricks and
hints from those how have successfully overcome the troubles of making yeast
nuclear extracts?

Thanks in advance                J.Gervais  Universite LAVAL Quebec

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