zymolyase stock solution
Malcolm Gardner
Dear yeast workers,
I am just starting to work with YAC libraries and I would like to know what
is the best way to prepare stock solutions of zymolyase 100T that could be
stored frozen.
None of the protocol books I have read describe how to prepare stock
solutions. Is the enzyme stable in aqueous solution?
Thanks very much for your help.
Malcolm J. Gardner, Ph.D.
The Institute for Genomic Research
9712 Medical Center Drive
Rockville, MD 20850
Phone: (301)-838-3519
FAX: (301)-838-0208
email: gar...@tigr.org
Namjin Chung
>Subject: Re: zymolyase stock solution
>Sent: 3-7-97 8:24p
>Received: 3-7-97 5:20p
>From: LEIB...@ocelot.rutgers.edu
>To: ye...@net.bio.net
>
>In my hands, I have found aqueous solutions of zymolyase not to be terribly
>stable, and are best made up fresh, stored in ice briefly, and discarded
>at the
>end of each day. I would also be interested if anyone knows how to store
>this.
>
>Mike Leibowitz
>UMDNJ-Robert Wood Johnson Medical School
At least for routine use for tetrad dissection, I have just dissolved
Zymolyase 20T (ICN, 2mg/mL) in water, filtered it, and used it
repeatedly. It seemed that it kept activity as long as it was stored in
a refrigerator. Cheers!
___________________________________
Namjin Chung
Rm. 213 MSRB, Box 3345
Program in Molecular Cancer Biology
Duke University Medical Center
Durham, NC 27710
n...@acpub.duke.edu
___________________________________
x...@cornell.edu
Zymolyase doesn't dissolve too well in pure water-- I have to use 1M
sorbitol or sucrose to dissolve it up. I usually make a 2mg/ml stock
solution in 1M sorbitol and freeze aliquots. The sorbitol probably also
helps protect against damage during freezing. I don't know exactly how
much activity gets lost with freezing and thawing, but I've never had a
problem even after several freeze/ thaws.
-Daniel
Ricardo Azpiroz
On 7 Mar 1997 LEIB...@ocelot.rutgers.edu wrote:
In my hands, I have found aqueous solutions of zymolyase not to be terribly
stable, and are best made up fresh, stored in ice briefly, and discarded at the
end of each day. I would also be interested if anyone knows how to store this.
Mike Leibowitz
UMDNJ-Robert Wood Johnson Medical School
I have found that solutions in 1M sorbitol, buffered with citrate at pH
5.8, (I can't recall the citrate conc.) are VERY stable and can be frozen
and thawed repeatedly.
----------------------
Ricardo Azpiroz
Department of Biochemistry
Life Sciences South
University of Arizona
Tucson AZ 85721-0106
Nathan W. Bays
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I use zymolyase in SEC buffer and store it at -20=B0C. It apparently
retains activity even after multiple freeze/thaws. Of course, I
immediately freeze any remaining solution after I use it.
SEC Buffer (100 ml stock)
18.2 g Sorbitol
2.94g Sodium Citrate
12 ml 0.5 M EDTA
adjust to pH 7 with HCl.
I add zymolyase to 5 mg/ml and filter sterilize solution.
Avoid nitrocellulose filter.
Hope this helps,
Nate
---------------------------------------------------------------
Nathan W. Bays
Ph.D. Student, Department of Biology
University of California, San Diego
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<X-FONTSIZE><PARAM>12</PARAM><FONTFAMILY><PARAM>Palatino</PARAM>I use
zymolyase in SEC buffer and store it at -20=B0C. It apparently retains
activity even after multiple freeze/thaws. Of course, I immediately
freeze any remaining solution after I use it.
SEC Buffer (100 ml stock)
18.2 g Sorbitol
2.94g Sodium Citrate
12 ml 0.5 M EDTA
adjust to pH 7 with HCl.
I add zymolyase to 5 mg/ml and filter sterilize solution.
Avoid nitrocellulose filter.
Hope this helps,
Nate
---------------------------------------------------------------
Nathan W. Bays
Ph.D. Student, Department of Biology
University of California, San Diego
</FONTFAMILY></X-FONTSIZE>
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