Need Info on Zymolyase for Tetrad Analysis

Beth A. Montelone bethmont at
Tue Jan 16 13:42:23 EST 1996

We use the really crude yucky brown glusulase which I think came from Sigma 
originally.  We dilute this 1 to 10 in sterile water and use the dilution for a
month or so until it loses activity.  A suspension of the sporulated mixture from
a plate is made in 0.2 M NaPO4, pH 8.0 or sterile water and glusulase added to
a final dilution of anywhere from 1 to 30 to 1 to 200 depending on the strain.  
Times for incubation at RT are 3 to 30 minutes, again depending on the strain.
You will have to determine the optimal conditions empirically for your strain, I'm 
afraid.  If nothing is digested, go longer times or add more glusulase.  If 
everything dies, go shorter or less glusulase.  We then make a streak using a loop 
from the suspension onto a plate or slab and have at it.

Good luck!

Beth Montelone
Kansas State University
bethmont at

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