<html><head><meta http-equiv="content-type" content="text/html; charset=us-ascii"></head><body style="overflow-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;"><div></div><br><div><br><blockquote type="cite"><div>On Jan 23, 2024, at 18:08, Zannati Zaoti <z.zaoti@uq.edu.au> wrote:</div><br class="Apple-interchange-newline"><div><meta charset="UTF-8"><div class="WordSection1" style="page: WordSection1; caret-color: rgb(0, 0, 0); font-family: Helvetica; font-size: 14px; font-style: normal; font-variant-caps: normal; font-weight: 400; letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; text-decoration: none;"><div style="margin: 0cm; font-size: 11pt; font-family: Aptos, sans-serif;"><span style="color: rgb(33, 33, 33);"><o:p> </o:p></span></div><div style="margin: 0cm; font-size: 11pt; font-family: Aptos, sans-serif;"><span style="color: rgb(33, 33, 33);">Dear all,<o:p></o:p></span></div><div style="margin: 0cm; font-size: 11pt; font-family: Aptos, sans-serif;"><span style="color: rgb(33, 33, 33);"> <o:p></o:p></span></div><div style="margin: 0cm; font-size: 11pt; font-family: Aptos, sans-serif;"><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.<o:p></o:p></span></div><div style="margin: 0cm; font-size: 11pt; font-family: Aptos, sans-serif;"><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.<o:p></o:p></span></div><div style="margin: 0cm; font-size: 11pt; font-family: Aptos, sans-serif;"><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. <o:p></o:p></span></div><div style="margin: 0cm; font-size: 11pt; font-family: Aptos, sans-serif;"><span style="color: rgb(33, 33, 33);">Any advice or comment would be highly appreciated. Thank you!<o:p></o:p></span></div><div style="margin: 0cm; font-size: 11pt; font-family: Aptos, sans-serif;"><span style="color: rgb(33, 33, 33);"> <o:p></o:p></span></div><div style="margin: 0cm; font-size: 11pt; font-family: Aptos, sans-serif;"><span style="color: rgb(33, 33, 33);"> <o:p></o:p></span></div><div style="margin: 0cm; font-size: 11pt; font-family: Aptos, sans-serif;"><span style="color: rgb(33, 33, 33);">Regards<o:p></o:p></span></div><div style="margin: 0cm; font-size: 11pt; font-family: Aptos, sans-serif;"><span style="color: rgb(33, 33, 33);">Zaoti <o:p></o:p></span></div><div style="margin: 0cm; font-size: 11pt; font-family: Aptos, sans-serif;"><o:p> </o:p></div><div style="margin: 0cm; font-size: 11pt; font-family: Aptos, sans-serif;"><o:p> </o:p></div></div><span style="caret-color: rgb(0, 0, 0); font-family: Helvetica; font-size: 14px; font-style: normal; font-variant-caps: normal; font-weight: 400; letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; text-decoration: none; float: none; display: inline !important;">_______________________________________________</span><br style="caret-color: rgb(0, 0, 0); font-family: Helvetica; font-size: 14px; font-style: normal; font-variant-caps: normal; font-weight: 400; letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; text-decoration: none;"><span style="caret-color: rgb(0, 0, 0); font-family: Helvetica; font-size: 14px; font-style: normal; font-variant-caps: normal; font-weight: 400; letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; text-decoration: none; float: none; display: inline !important;">3dem mailing list</span><br style="caret-color: rgb(0, 0, 0); font-family: Helvetica; font-size: 14px; font-style: normal; font-variant-caps: normal; font-weight: 400; letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; text-decoration: none;"><a href="mailto:3dem@ncmir.ucsd.edu" style="font-family: Helvetica; font-size: 14px; font-style: normal; font-variant-caps: normal; font-weight: 400; letter-spacing: normal; orphans: auto; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: auto; word-spacing: 0px; -webkit-text-stroke-width: 0px;">3dem@ncmir.ucsd.edu</a><br style="caret-color: rgb(0, 0, 0); font-family: Helvetica; font-size: 14px; font-style: normal; font-variant-caps: normal; font-weight: 400; letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; text-decoration: none;"><a href="https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem" style="font-family: Helvetica; font-size: 14px; font-style: normal; font-variant-caps: normal; font-weight: 400; letter-spacing: normal; orphans: auto; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: auto; word-spacing: 0px; -webkit-text-stroke-width: 0px;">https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem</a></div></blockquote></div><br></body></html>