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



More information about the Yeast mailing list