Title: Re: [ccp4bb]: spherulites
Dear Jacob,
 
What additives do you suggest that actually will promote crystallization of DNA and RNA complexes and get me over this hurdle to the microcrystal stage?
 
I am trying of course the usual cast of suspects; divalent cations like Mg, Mn, Zn, Ca etc, various anions and spermine etc
 
From all of the responses that I have got, it looks like changing the oligo lengths may be the best answer, if the RNA refuses to crystallize well.
 
 
Many thanks
 
Ray
 
Ray Brown
Department of Molecular and Cell Biology
University of Connecticut

From: Jacob Corn [mailto:[EMAIL PROTECTED]
Sent: Tue 9/5/2006 12:22 PM
To: Brown,Raymond (BIDMC - Experimental Medicine)
Subject: Re: [ccp4bb]: spherulites

Hi Ray,
I was able to turn spherulites of a DNA-protein complex into finicky plates
(5-10 x 200 x 200 um) that diffract (very anisotropically) to 3.0A. and I've so
far been able to collect a mostly-complete native set and solve it by MR (with
density for the DNA visible). So while this may not be water into wine, think of
it as water into grape juice.

Anyway, what I started with was additive seeding. This helped me turn spherulite
fuzzballs into single itsy-needles. I then did several rounds of additive
screening in an attempt to make the xtals larger and more defined. I finished up
by pursuing many different oligo lengths until I found one that yielded the
best-diffracting plates.

Jacob

[EMAIL PROTECTED] wrote:
> Hi all,

> I have now progressed from phase separation to growing many nice
> spherulites, after many months of trying to crystallize some RNA-protein
> complexes. Has anybody out there been able to turn water into wine?

> If you have had any experience of successfully changing spherulites into
> single crystals with PEG, MPD, HD etc, then I would really like to hear
> about your success story.

> Any practical suggestions or tips are welcome?

> Cheers,


> Ray


> [EMAIL PROTECTED] <mailto:[EMAIL PROTECTED]>


--
Jacob Corn
The Berger Lab
UC Berkeley - Molecular and Cell Biology
[EMAIL PROTECTED]
phone: 510-643-8893
fax: 510-643-9290

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