That's a really old paper. You can purchase the lysozyme from Hampton
Research and it's fine. The recipe is available from the Hampton Research
page:
http://hamptonresearch.com/product_detail.aspx?cid=28&sid=173&pid=524
Grow them a low temp and you can stop them when they are the right size.
I favor that over room temp. They grow fast and large, but don't give a
good R(merge) as when grown at a lower concentration and slower.
Bernie
On Tue, July 26, 2011 5:09 pm, [log in to unmask] wrote:
>
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> James,
>
> I would have a look at the paper by Judge et al:
>
> http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1300446/pdf/10465769.pdf
>
>
> Specifically, in this paper you will find that the crystallization
> behavior of lysozyme changes drastically with pH. At the time the paper
> wasn't really written to manipulate for small crystal size, but looking
> back at the paper (specifically Fig 5), it appears that you can read the
> conditions that will give you crystals around the size you want.
>
> Not re-reading the paper, quoting from memory (which we all think is
> better than it really is), it is important to use good quality lysozyme to
> get reproducible results. Good quality probably means freshly purified
> from fresh (farm-acquired) eggs. I am not kidding you, it makes a big
> difference. Also, I am going out on a limb to say (I know you know this)
> that the buffer preparation method matters a lot. Taking sodium acetate
> solution and pH-ing it with HCl will give very different results from
> taking acetic acid and pH-ing it with NaOH (because the ionic strength of
> the buffer is not the same). Lysozyme crystallizes so easily that we tend
> to forget tedious details.
>
> Hope this helps. This paper will probably give you some ideas in the right
> direction.
>
> Mark van der Woerd
>
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