<div dir="ltr"><div>Perhaps ultraviolet light induced crosslinking would be worth a shot.<br></div><div><br></div><div>Best,</div><div>-da<br></div></div><br><div class="gmail_quote"><div dir="ltr" class="gmail_attr">On Tue, Jan 23, 2024 at 6:08 PM Zannati Zaoti <<a href="mailto:z.zaoti@uq.edu.au">z.zaoti@uq.edu.au</a>> wrote:<br></div><blockquote class="gmail_quote" style="margin:0px 0px 0px 0.8ex;border-left:1px solid rgb(204,204,204);padding-left:1ex"><div class="msg5077699108472731428">
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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
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<p class="MsoNormal"><span style="color:rgb(33,33,33)"><u></u> <u></u></span></p>
<p class="MsoNormal"><span style="color:rgb(33,33,33)">Dear all,<u></u><u></u></span></p>
<p class="MsoNormal"><span style="color:rgb(33,33,33)"> <u></u><u></u></span></p>
<p class="MsoNormal"><span style="color:rgb(33,33,33)">I am looking for advice from experienced individuals who have performed cross-linking of DNA-bound protein complexes for cryo-EM sample preparation.<u></u><u></u></span></p>
<p class="MsoNormal"><span style="color:rgb(33,33,33)">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.<u></u><u></u></span></p>
<p class="MsoNormal"><span style="color:rgb(33,33,33)">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. <u></u><u></u></span></p>
<p class="MsoNormal"><span style="color:rgb(33,33,33)">Any advice or comment would be highly appreciated. Thank you!<u></u><u></u></span></p>
<p class="MsoNormal"><span style="color:rgb(33,33,33)"> <u></u><u></u></span></p>
<p class="MsoNormal"><span style="color:rgb(33,33,33)"> <u></u><u></u></span></p>
<p class="MsoNormal"><span style="color:rgb(33,33,33)">Regards<u></u><u></u></span></p>
<p class="MsoNormal"><span style="color:rgb(33,33,33)">Zaoti <u></u><u></u></span></p>
<p class="MsoNormal"><u></u> <u></u></p>
<p class="MsoNormal"><u></u> <u></u></p>
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