Lysozyme stock - (Mar/04/2009 )
I am making Lysozyme stock (100mg/ml) that I will use in lysis buffer.
Just a few questions...
I guess people suspend the enzyme in different liquid, but sterilized water should be just fine?--> I wonder if lysozme is more stable in other kinds of buffer?
Also, since I cant autoclave this, is it best to filter sterilize lysozyme stock or no need to sterilize?
How do you guys sterilize Triton X-100? Filter? or Autoclave?
I prepare lysozyme stock in TE buffer. Lysozyme is most active in TE at pH 8-9. You don't need to sterilize lysozyme stock as it would not touch your cultures.
What about just Tris 8-9? Most enzyme i think require Mg2+ to function properly but i don't know if this lysozyme is one of it. However another thing about this is .. i really doubt the stock pH matters that much or not because you're not going to add a whole lot of it during ur genomic extraction / plasmid extraction prep. It's the resuspension buffer that might have the final say.
Thanks for the replies.
I am just curious, but the reason you do not have to filter-sterilize the lysozyme is because you use it to lyse the cells and nothing can grown in it?
(There is no harm in sterilizing it thought right?) I am a freak when it comes to sterilizing things.
Once the lysozyme stock is made, do you aliqoute small volumes and throw away unused portion after thawing once?
I am not sure if lysozyme is freeze-thaw sensitive or not.
Ecoli0157 on Mar 4 2009, 07:06 PM said:
No need to sterilize Triton X-100.
Nothing can grow in there (cells will be lysed).
Just prepare fresh solution.