Hi all. Just wanted to add a bit of doubt to the topic. Is it possible that
they are salt crystals? I don’t know, but some spots are too intense and far
from each other to me. But maybe I just need better glasses.
Best wishes
Rafael Marques da Silva
PhD Student – Structural Biology
University of Leicester
Mestre em Física Biomolecular
Universidade de São Paulo
Bacharel em Ciências Biológicas
Universidade Federal de São Carlos
phone: +55 16 99766-0021
"A sorte acompanha uma mente bem treinada"
________________________________________________
De:
[email protected]<mailto:[email protected]>
Enviado:sexta-feira, 24 de novembro de 2023 09:00
Para: [email protected]<mailto:[email protected]>
Assunto: Re: [ccp4bb] What could these crystals be?
I wanted to thank everyone for their suggestions and ideas regarding the
eyeball shaped crystals that I got at the beginning of the month.
I can confirm that these crystals do indeed contain protein and DNA which is
good news.
I have tried a number of different buffers and salts since then. I have also
tried seeding but nothing I have tried has changed the morphology of the
crystals. They remain thin, flat and eyeball/pumpkin seed shaped.
It is not difficult to make the crystals, they form quite easily under quite a
few conditions. The difficulty is in the diffraction. By changing the buffer
conditions we can now see some very weak diffraction (previously there was no
diffraction at all).
Are there any suggestions as to how to improve diffraction of these crystals? I
did try different cryoprotectants, parabar, glycerol, PEG but no difference. I
think perhaps the problem is heterogeneity considering my sample contains both
protein and DNA.
Any suggestions or thoughts are welcome.
Thank you
Careina
On Wednesday, November 8, 2023 at 06:18:46 PM GMT+2, [email protected]
<[email protected]> wrote:
The reservior solution is 0.2 M NaCl2, 0.1 M HEPES pH 7.5, and 25% 3350 PEG
Protein buffer is 300 mM NaCl, 20 mM HEPES pH 7.5, 3mM TCEP
The drop contains 1.5ul protein-DNA complex and 1.5 ul reservior solution
On Wednesday, November 8, 2023 at 05:12:25 PM GMT+2,
[email protected] <[email protected]> wrote:
Hi Careina,
Without knowing what's in your protein buffer or crystallisation condition it's
hard to comment.
Best wishes,
Steve
Dr Stephen Carr
Research Complex at Harwell (RCaH)
Rutherford Appleton Laboratory
Harwell Oxford
Didcot
Oxon OX11 0FA
United Kingdom
Email [email protected]
tel 01235 567717
From: CCP4 bulletin board <[email protected]> on behalf of
[email protected] <[email protected]>
Sent: 08 November 2023 15:00
To: ccp4bb <[email protected]>
Subject: [ccp4bb] What could these crystals be?
Hi all
We have been trying with no success to crystalize a protein. Recently we got
these strange shape "crystals". They are hard and flat but they do not diffract
at all. Any ideas as to what could cause this?
Careina
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