Hi Careina.
The top two things that come to my mind is that you need to screen for
different crystallization conditions and do some seeding. If your Crystal
conditions does not diffract at all, probably it won’t. Have you tried to
optimize your condintion changing the pH or precipitant concentration?
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"
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De: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> em nome de
careinaedgo...@yahoo.com <000002531c126adf-dmarc-requ...@jiscmail.ac.uk>
Enviado: Thursday, February 8, 2024 8:25:25 AM
Para: CCP4BB@JISCMAIL.AC.UK <CCP4BB@JISCMAIL.AC.UK>
Assunto: [ccp4bb] Crystals with DNA
Hello all.
I am struggling to get defracting crystals with a protein DNA complex. The
crystals are plentiful but they do not diffract. I am going back to the grind
stone and relookong at my DNA sequence.
Is there any wisdom you could give me with regards to what works best with DNA
in crystals?
>From my reading it seems if the length is a multiple of 7 (for B DNA) and
>blunt ended, it will stretch over the length of the crystal and improve
>crystalisability. But if you want crystals that diffract better, you will need
>to play with length and even making it only one base longer or shorter can
>make a difference, even changing the morphology of the crystal? Longer is
>better than shorter, and overhangs are good for improving diffraction?
>Presumably because they stabilize contacts? It is expensive to synthesize a
>while bunch of sequences so I need to be strategic in my choice. Would
>appreciate any advice.
Thank you
Careina.
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