[3dem] DNA-protein cross-linker

[Daniel Asarnow]

Perhaps ultraviolet light induced crosslinking would be worth a shot.

Best,
-da

On Tue, Jan 23, 2024 at 6:08 PM Zannati Zaoti <z.zaoti at uq.edu.au> wrote:

> Dear all, I am looking for advice from experienced individuals who have
> performed cross-linking of DNA-bound protein complexes for cryo-EM sample
> preparation. My work involves a protein that can bind to dsDNA and form
> filaments; however, these
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> Dear all,
>
>
>
> I am looking for advice from experienced individuals who have performed
> cross-linking of DNA-bound protein complexes for cryo-EM sample preparation.
>
> My work involves a protein that can bind to dsDNA and form filaments;
> however, these filaments are flexible and tend to fall apart during grid
> preparation. As a result, I cannot find any filaments during the cryoEM
> screening.
>
> Thereafter, I have tried using glutaraldehyde and formaldehyde as
> cross-linkers to stabilize the filaments, but they have made the filaments
> even more unstable. At present, I am experimenting with DMS, but it seems
> that DMS neither makes the filament unstable nor stable enough for
> cryo-conditions.
>
> Any advice or comment would be highly appreciated. Thank you!
>
>
>
>
>
> Regards
>
> Zaoti
>
>
>
>
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