4 replies to this topic

[Ecoli0157]

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?

Thanks,

Edited by Ecoli0157, 04 March 2009 - 12:54 PM.

[AquaPlasmid]

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.

[hanming86]

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.

[Ecoli0157]

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.

Edited by Ecoli0157, 09 March 2009 - 11:26 AM.

[molgen]

Ecoli0157, on Mar 4 2009, 07:06 PM, said:

How do you guys sterilize Triton X-100? Filter? or Autoclave?

Thanks,

No need to sterilize Triton X-100.
Nothing can grow in there (cells will be lysed).
Just prepare fresh solution.