lyticase treatment

Astrid Helfant ahelfant at mail.colgate.edu
Wed Aug 16 15:23:23 EST 2000


> My yeast cells have been "crashing" upon:
> - 60 min. formaldehyde fixation at RT in the presence of 50 mM KPO4 and 50
> mM MgCl2
> - 15-30 min. lyticase treatment at 37'C in the presence of 50 mM KPO4 and
> 50 mM MgCl2 and 0.5 M sorbitol
> - incubation in 1x PBS at RT
> 
> The cells basically look like they've been under severe osmotic stress
> which results in their "crashing."
> I ordered and made all new solutions and am now running out of options.
> Does anybody have any suggestions? Would perhaps 
> treatment with gluculase or zymolase instead of lyticase prevent this from
> happening? Perhaps a different fixation technique? 
> Perhaps adding more salt to my buffers?
> 
> Astrid Helfant
> Colgate University
> Hamilton New York
> e-mail: ahelfant at mail.colgate.edu
> 


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