Hi Mayer and Jason,

Thanks for your quick reply! I read from Hampton that for PEG smaller than
5K, directly increasing its concentration is a way of making cryo. But
you're right, glycerol or PEG400 will be a good way and might be easier.
I'll try that first.

Thanks a lot!

Best,
Xiao

2014-10-13 17:59 GMT-07:00 Mayer, Mark (NIH/NICHD) [E] <[email protected]>
:

> Try either glycerol or PEG 400.
> Increasing concentration of 'high' MW PEG is not a good strategy.
>
> ________________________________________
> From: Xiao Xiao [[email protected]]
> Sent: Monday, October 13, 2014 8:51 PM
> To: [email protected]
> Subject: [ccp4bb] A quick question about making PEG cryo-protectant
>
> Hi,
>
> Since my crystallization condition contains PEG, I am trying to use PEG as
> one of my cryo for testing. For example, one crystallization condition is
> 0.2M Na2PO4, 20%PEG3350. I want to make 0.2M Na2PO4 and 40% PEG3350 as
> cryoprotectant. When I added 200ul salt from 1M stock, and 800ul PEG from
> 50% stock, mixed them, seems they stayed as emulsion (confirmed by
> microscope), no matter how I pipette or vortex them. I tried to incubate it
> at 40degree, seems it got worse.
>
> I am now trying to re-make the cryo from solid reagent, not solution
> stock. Will this be helpful? Or is there any other trick?
>
> I will really appreciate!
>
> Best,
> Xiao
>

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