[3dem] DNA-protein cross-linker

[Zannati Zaoti]

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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