enzyme lysis

S L Forsburg forsburg at nospamsalk.edu
Mon Dec 1 19:21:01 EST 1997


> From: Celedonio =?iso-8859-1?Q?Gonz=E1lez?= <cglez at ull.es>

> Does anybody know of a method to prepare nuclear extracts from yeast
> without the use of Zymolyase?
> 
> I need to prepare nuclear extracts from Hansenula polymorpha, a yeast
> whose wall breaks very poorly with Zymolyase. A protocol that uses
> glass
> beads to break the cells will be wonderful

Different enzymes break different linkages in the cell wall, and 
different yeasts have different carbohydrate structures.  Have
you tried alternatives, such as Novozyme (favored for pombe),
or glusalase, or a cocktail of multiple enzymes?  I'm concerned
that glass bead breakage will disrupt your nuclei as well as
your cell wall.



-- 
-susan
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Molecular Biology and Virology Lab          
The Salk Institute, La Jolla CA 
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