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<p class="MsoNormal"><span style="font-size:11.0pt">Hello Daniel,</span></p>
<p class="MsoNormal"><span style="font-size:11.0pt"> </span></p>
<p class="MsoNormal"><span style="font-size:11.0pt">Thank you for your response.<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt">I don’t know much about it if UV-induced crosslinkers may cause any damaging effects on the sample. <o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt">Regards<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:11.0pt">Zaoti </span></p>
<p class="MsoNormal"><span style="font-size:11.0pt"> </span></p>
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<p class="MsoNormal" style="margin-bottom:12.0pt"><b><span style="color:black">From:
</span></b><span style="color:black">Daniel Asarnow <asarnow@msg.ucsf.edu><br>
<b>Date: </b>Wednesday, 24 January 2024 at 12:21</span><span style="font-family:"Arial",sans-serif;color:black"> </span><span style="color:black">pm<br>
<b>To: </b>Zannati Zaoti <z.zaoti@uq.edu.au><br>
<b>Cc: </b>3dem@ncmir.ucsd.edu <3dem@ncmir.ucsd.edu>, 3dem-request@ncmir.ucsd.edu <3dem-request@ncmir.ucsd.edu><br>
<b>Subject: </b>Re: [3dem] DNA-protein cross-linker</span></p>
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<p class="MsoNormal">Perhaps ultraviolet light induced crosslinking would be worth a shot.</p>
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<p class="MsoNormal"> </p>
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<p class="MsoNormal">Best,</p>
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<p class="MsoNormal">-da</p>
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<p class="MsoNormal"> </p>
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<p class="MsoNormal">On Tue, Jan 23, 2024 at 6:08<span style="font-family:"Arial",sans-serif"> </span>PM Zannati Zaoti <<a href="mailto:z.zaoti@uq.edu.au">z.zaoti@uq.edu.au</a>> wrote:</p>
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<p class="MsoNormal"><span style="font-size:1.0pt;color:white">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 </span></p>
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<p class="MsoNormal"><span style="color:#212121"> </span></p>
<p class="MsoNormal"><span style="color:#212121">Dear all,</span></p>
<p class="MsoNormal"><span style="color:#212121"> </span></p>
<p class="MsoNormal"><span style="color:#212121">I am looking for advice from experienced individuals who have performed cross-linking of DNA-bound protein complexes for cryo-EM sample preparation.</span></p>
<p class="MsoNormal"><span style="color:#212121">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.</span></p>
<p class="MsoNormal"><span style="color:#212121">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. </span></p>
<p class="MsoNormal"><span style="color:#212121">Any advice or comment would be highly appreciated. Thank you!</span></p>
<p class="MsoNormal"><span style="color:#212121"> </span></p>
<p class="MsoNormal"><span style="color:#212121"> </span></p>
<p class="MsoNormal"><span style="color:#212121">Regards</span></p>
<p class="MsoNormal"><span style="color:#212121">Zaoti </span></p>
<p class="MsoNormal"> </p>
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