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I don't have too much experience with detergents, but when I have tried I've had most success with UA, so no escaping those pesky uranyl salts completely. </div>
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<div id="divRplyFwdMsg" dir="ltr"><font face="Calibri, sans-serif" style="font-size:11pt" color="#000000"><b>From:</b> Craig Yoshioka <yoshiokc@ohsu.edu><br>
<b>Sent:</b> 21 October 2021 16:16<br>
<b>To:</b> Lesley Calder <Lesley.Calder@crick.ac.uk><br>
<b>Cc:</b> 3dem@ncmir.ucsd.edu <3dem@ncmir.ucsd.edu><br>
<b>Subject:</b> Re: [3dem] Uranyl acetate alternative</font>
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<span style="font-size:10.0pt; color:red"><b>External Sender:</span></b> <span style="font-size:10.0pt; color:black">
Use caution.</span></div>
<div>This is great, thanks! Out of curiosity do you have a goto alternative if looking at samples with detergents?<br class="">
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<div class="">On Oct 21, 2021, at 5:58 AM, Lesley Calder <<a href="mailto:Lesley.Calder@crick.ac.uk" class="">Lesley.Calder@crick.ac.uk</a>> wrote:</div>
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It seems like quite a few of you are interested in the SST preparation protocol, so here it is attached.</div>
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Lesley</div>
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<font face="Calibri, sans-serif" class="" style="font-size:11pt"><b class="">From:</b><span class="x_Apple-converted-space"> </span>Metskas, Lauren Ann <<a href="mailto:lmetskas@purdue.edu" class="">lmetskas@purdue.edu</a>><br class="">
<b class="">Sent:</b><span class="x_Apple-converted-space"> </span>21 October 2021 12:12<br class="">
<b class="">To:</b><span class="x_Apple-converted-space"> </span>Lesley Calder <<a href="mailto:Lesley.Calder@crick.ac.uk" class="">Lesley.Calder@crick.ac.uk</a>><br class="">
<b class="">Subject:</b><span class="x_Apple-converted-space"> </span>Re: [3dem] [EXTERNAL] Uranyl acetate alternative</font>
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<span class="" style="font-size:10pt; color:red"><b class="">External Sender:</b></span><span class="x_Apple-converted-space"> </span><span class="" style="font-size:10pt">Use caution.</span></div>
<div class="">Hello,
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<div class="">I’d love your recipe for the conversion to the salt form if you’re willing to share it! I’m just starting up a new lab at Purdue University, and have been thinking about switching to a non-radioactive stain.</div>
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<div class="">Thank you! Best,</div>
<div class="">Lauren Ann</div>
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Lauren Ann Metskas, Ph.D.</div>
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Assistant Professor<br class="">
Department of Biological Sciences<br class="">
Joint Appointment in Department of Chemistry</div>
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Purdue University</div>
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Hockmeyer Hall (HOCK) Room 231<br class="">
240 S Martin Jischke Dr.<br class="">
West Lafayette, IN 47907<br class="">
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(Office) (765) 49-44909</div>
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<a href="mailto:metskas@purdue.edu" class="">metskas@purdue.edu</a></div>
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<blockquote type="cite" class="">
<div class="">On Oct 21, 2021, at 5:15 AM, Lesley Calder <<a href="mailto:Lesley.Calder@crick.ac.uk" class="">Lesley.Calder@crick.ac.uk</a>> wrote:</div>
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In 35 years of negative staining my go to stain is sodium silicotungstate. It provides good contrast has a fine grain, and therefore reveals details not obvious with uranyl salts and is great for really small molecules, it has none of the problems of radioactivity
for handling and disposal. I use it with all the conventional buffers including PBS, but it's of no use when detergent is present. </div>
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UK suppliers sell the acid form (not sure about anywhere else), and to achieve the best affects you need to neutralise your solution. I prefer to prepare a batch of the salt from the acid solution and precipitate it out to keep the solid for later use ... it
keeps for years that way. I then prepare a 1% solution of the salt form, which also keeps for many months. </div>
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If anyone would like the recipe for the conversion to the salt form, I'd be happy to supply it.</div>
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Lesley Calder</div>
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<span class="" style="font-size:10pt">The Francis Crick Institute</span><br class="">
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<span class="" style="margin:0px; font-size:10pt">1 Midland Road</span></div>
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<span class="" style="margin:0px; font-size:10pt">London NW1 1AT</span></div>
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<font face="Calibri, sans-serif" class="" style="font-size:11pt"><b class="">From:</b><span class="x_x_Apple-converted-space"> </span>3dem <<a href="mailto:3dem-bounces@ncmir.ucsd.edu" class="" style="color:purple; text-decoration:underline">3dem-bounces@ncmir.ucsd.edu</a>>
on behalf of Riedel, Dietmar <<a href="mailto:driedel@gwdg.de" class="" style="color:purple; text-decoration:underline">driedel@gwdg.de</a>><br class="">
<b class="">Sent:</b><span class="x_x_Apple-converted-space"> </span>20 October 2021 14:32<br class="">
<b class="">To:</b><span class="x_x_Apple-converted-space"> </span>Sharon G Wolf <<a href="mailto:Sharon.Wolf@weizmann.ac.il" class="" style="color:purple; text-decoration:underline">Sharon.Wolf@weizmann.ac.il</a>><br class="">
<b class="">Cc:</b><span class="x_x_Apple-converted-space"> </span><a href="mailto:3dem@ncmir.ucsd.edu" class="" style="color:purple; text-decoration:underline">3dem@ncmir.ucsd.edu</a><span class="x_x_Apple-converted-space"> </span><<a href="mailto:3dem@ncmir.ucsd.edu" class="" style="color:purple; text-decoration:underline">3dem@ncmir.ucsd.edu</a>><br class="">
<b class="">Subject:</b><span class="x_x_Apple-converted-space"> </span>Re: [3dem] [EXTERNAL] Uranyl acetate alternative</font>
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<span class="" style="font-size:10pt; color:red"><b class="">External Sender:</b></span><span class="x_x_Apple-converted-space"> </span><span class="" style="font-size:10pt">Use caution.</span></div>
<div class="">
<div dir="ltr" class="">I use NanoVan or NanoW. NanoVan is great for negativ stain if phosphates omit the use of Uran stains. But it is a vanadium stain and therefore much weaker than Uran salt. I keep it in the fridge for years. </div>
<div dir="ltr" class="">Dietmar <br class="">
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<div dir="ltr" class="">Mobil gesendet</div>
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<blockquote type="cite" class="">Am 20.10.2021 um 15:07 schrieb Sharon G Wolf <<a href="mailto:Sharon.Wolf@weizmann.ac.il" class="">Sharon.Wolf@weizmann.ac.il</a>>:<br class="">
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<span class="" style="font-size:11pt; font-family:Calibri,sans-serif; color:rgb(31,73,125)">I have used sodium silicotungstate on a variety of samples. Also phosphotungstic acid is a classic.</span></div>
<div class="" style="margin:0in 0in 0.0001pt; font-size:12pt; font-family:"Times New Roman",serif">
<span class="" style="font-size:11pt; font-family:Calibri,sans-serif; color:rgb(31,73,125)">Also, like Zuben, I have used ammonium molybdate and methylamine tungstate. Both need to be made fresh. They can’t sit on the shelf for a long time like UAc can.<br class="">
None of them has as much contrast as UAc, except for the sodium silicotungstate, which can be outstanding. But it can cause some complexes to fall apart.</span></div>
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<span class="" style="font-size:11pt; font-family:Calibri,sans-serif; color:rgb(31,73,125)">Sharon</span></div>
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<span class="" style="font-size:11pt; font-family:Calibri,sans-serif; color:rgb(31,73,125)"> </span></p>
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<b class=""><span class="" style="font-size:11pt; font-family:Calibri,sans-serif">From:</span></b><span class="" style="font-size:11pt; font-family:Calibri,sans-serif"><span class="x_x_Apple-converted-space"> </span>3dem <<a href="mailto:3dem-bounces@ncmir.ucsd.edu" class="">3dem-bounces@ncmir.ucsd.edu</a>><span class="x_x_Apple-converted-space"> </span><b class="">On
Behalf Of<span class="x_x_Apple-converted-space"> </span></b>Brown, Zuben<br class="">
<b class="">Sent:</b><span class="x_x_Apple-converted-space"> </span>Wednesday, 20 October, 2021 15:41<br class="">
<b class="">To:</b><span class="x_x_Apple-converted-space"> </span>Denis Chrétien <<a href="mailto:denis.chretien@univ-rennes1.fr" class="">denis.chretien@univ-rennes1.fr</a>>;<span class="x_Apple-converted-space"> </span><a href="mailto:3dem@ncmir.ucsd.edu" class="">3dem@ncmir.ucsd.edu</a><br class="">
<b class="">Subject:</b><span class="x_x_Apple-converted-space"> </span>Re: [3dem] [EXTERNAL] Uranyl acetate alternative</span></div>
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<span class="" style="font-family:Calibri,sans-serif">Hi Denis,</span></div>
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<span class="" style="font-family:Calibri,sans-serif">I've tried ammonium molybdate and methylamine tungstate with acceptable results on different membrane proteins.</span></div>
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<span class="" style="font-family:Calibri,sans-serif">Obviously, the contrast is reduced compared to uranyl acetate but the management of the waste materials is greatly simplified.</span></div>
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<span class="" style="font-family:Calibri,sans-serif">best wishes,</span></div>
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<span class="" style="font-family:Calibri,sans-serif">Zuben</span></div>
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<b class=""><span class="" style="font-size:11pt; font-family:Calibri,sans-serif">From:</span></b><span class="" style="font-size:11pt; font-family:Calibri,sans-serif"><span class="x_x_Apple-converted-space"> </span>3dem <<a href="mailto:3dem-bounces@ncmir.ucsd.edu" class="" style="color:purple; text-decoration:underline">3dem-bounces@ncmir.ucsd.edu</a>>
on behalf of Denis Chrétien <<a href="mailto:denis.chretien@univ-rennes1.fr" class="" style="color:purple; text-decoration:underline">denis.chretien@univ-rennes1.fr</a>><br class="">
<b class="">Sent:</b><span class="x_x_Apple-converted-space"> </span>Wednesday, October 20, 2021 3:35 AM<br class="">
<b class="">To:</b><span class="x_x_Apple-converted-space"> </span><a href="mailto:3dem@ncmir.ucsd.edu" class="" style="color:purple; text-decoration:underline">3dem@ncmir.ucsd.edu</a><span class="x_x_Apple-converted-space"> </span><<a href="mailto:3dem@ncmir.ucsd.edu" class="" style="color:purple; text-decoration:underline">3dem@ncmir.ucsd.edu</a>><br class="">
<b class="">Subject:</b><span class="x_x_Apple-converted-space"> </span>[EXTERNAL] [3dem] Uranyl acetate alternative</span></div>
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Dear all,</div>
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It becomes difficult to posses Uranyl Acetate salts. Hence we would like to have your feedback concerning alternatives for sections and negative staining. Along this line, we would appreciate your experience concerning this salt :</div>
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Other suggestions are welcomed!</div>
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Best,</div>
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Denis</div>
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<span class="">___________________________<br class="">
Université de Rennes 1<br class="">
UMR 6290, équipe TIPs<br class="">
Campus de Beaulieu, Bât. 13<br class="">
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France<br class="">
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Tel. +33 (0)2 23 23 67 64<br class="">
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<a href="https://urldefense.proofpoint.com/v2/url?u=https-3A__eur03.safelinks.protection.outlook.com_-3Furl-3Dhttps-253A-252F-252Furldefense.proofpoint.com-252Fv2-252Furl-253Fu-253Dhttps-2D3A-5F-5Feur03.safelinks.protection.outlook.com-5F-2D3Furl-2D3Dhttps-2D253A-2D252F-2D252Fmail.ncmir.ucsd.edu-2D252Fmailman-2D252Flistinfo-2D252F3dem-2D26data-2D3D04-2D257C01-2D257C-2D257Cb694a84aa2a8407b8fee08d99483a8d5-2D257C4eed7807ebad415aa7a99170947f4eae-2D257C0-2D257C1-2D257C637704115481237086-2D257CUnknown-2D257CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0-2D253D-2D257C3000-2D26sdata-2D3Dv3oz-2D252FoTiz6uAYcwVm9fTbNsnlsTjxl9qP4V1rEaqheE-2D253D-2D26reserved-2D3D0-2526d-253DDwMGaQ-2526c-253D-2D35OiAkTchMrZOngvJPOeA-2526r-253DL7-2DzyQ-2D04fFCMRqzLIOnx7H0exGZHwIQe-5FwMPuY600I-2526m-253DP1bCWoUI1BGBKRmq6cIUKsru5HLsgKAVcz8r0AigtoI-2526s-253D6km6Neve-5FpFVkSnynF50OuyXfHsa5yULLymFxp3vBmo-2526e-253D-26data-3D04-257C01-257C-257Cd583a9cb213e43961e6908d994a5c5b2-257C4eed7807ebad415aa7a99170947f4eae-257C0-257C1-257C637704262000906937-257CUnknown-257CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0-253D-257C3000-26sdata-3D7VPP-252B087aKANFufkHcb05MNGMAGhYKqM-252BrD-252BxyCVbPo-253D-26reserved-3D0&d=DwMGaQ&c=-35OiAkTchMrZOngvJPOeA&r=L7-zyQ-04fFCMRqzLIOnx7H0exGZHwIQe_wMPuY600I&m=tI8xjaUudfF6hGCXLS5V5WtJs6dKaA3BCqrcotjccN8&s=abBbHVc-cjq_uYraZNpinu7nt2kZVY4TDlWlqXT974o&e=" originalsrc="https://urldefense.proofpoint.com/v2/url?u=https-3A__eur03.safelinks.protection.outlook.com_-3Furl-3Dhttps-253A-252F-252Fmail.ncmir.ucsd.edu-252Fmailman-252Flistinfo-252F3dem-26data-3D04-257C01-257C-257Cb694a84aa2a8407b8fee08d99483a8d5-257C4eed7807ebad415aa7a99170947f4eae-257C0-257C1-257C637704115481237086-257CUnknown-257CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0-253D-257C3000-26sdata-3Dv3oz-252FoTiz6uAYcwVm9fTbNsnlsTjxl9qP4V1rEaqheE-253D-26reserved-3D0&d=DwMGaQ&c=-35OiAkTchMrZOngvJPOeA&r=L7-zyQ-04fFCMRqzLIOnx7H0exGZHwIQe_wMPuY600I&m=P1bCWoUI1BGBKRmq6cIUKsru5HLsgKAVcz8r0AigtoI&s=6km6Neve_pFVkSnynF50OuyXfHsa5yULLymFxp3vBmo&e=" shash="Dmqoz9LCvoe60aeqpKUsQlBpOHimczeKZUSbCd3PTjAjUIDzr3P+yf1gbKg2cK0MDYrCO+zBJWcBTEXSafPECrSCBsxL16ux/fuXglPmhtDNe5/4R5S7gxU+cfVtQveoQddX/MPMJQKJ9EjF1dYX0mQn37Xiz+pvIaaXAXs1BGA=" originalsrc="https://urldefense.proofpoint.com/v2/url?u=https-3A__eur03.safelinks.protection.outlook.com_-3Furl-3Dhttps-253A-252F-252Fmail.ncmir.ucsd.edu-252Fmailman-252Flistinfo-252F3dem-26data-3D04-257C01-257C-257Cb694a84aa2a8407b8fee08d99483a8d5-257C4eed7807ebad415aa7a99170947f4eae-257C0-257C1-257C637704115481237086-257CUnknown-257CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0-253D-257C3000-26sdata-3Dv3oz-252FoTiz6uAYcwVm9fTbNsnlsTjxl9qP4V1rEaqheE-253D-26reserved-3D0&d=DwMGaQ&c=-35OiAkTchMrZOngvJPOeA&r=L7-zyQ-04fFCMRqzLIOnx7H0exGZHwIQe_wMPuY600I&m=P1bCWoUI1BGBKRmq6cIUKsru5HLsgKAVcz8r0AigtoI&s=6km6Neve_pFVkSnynF50OuyXfHsa5yULLymFxp3vBmo&e=" shash="Dmqoz9LCvoe60aeqpKUsQlBpOHimczeKZUSbCd3PTjAjUIDzr3P+yf1gbKg2cK0MDYrCO+zBJWcBTEXSafPECrSCBsxL16ux/fuXglPmhtDNe5/4R5S7gxU+cfVtQveoQddX/MPMJQKJ9EjF1dYX0mQn37Xiz+pvIaaXAXs1BGA=" class="" style="color:purple; text-decoration:underline; font-family:Helvetica; font-size:12px; font-style:normal; font-variant-caps:normal; font-weight:normal; letter-spacing:normal; text-align:start; text-indent:0px; text-transform:none; white-space:normal; word-spacing:0px">https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem</a></div>
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The Francis Crick Institute Limited is a registered charity in England and Wales no. 1140062 and a company registered in England and Wales no. 06885462, with its registered office at 1 Midland Road London NW1 1AT</div>
<span id="x_cid:2F177273-383A-49C0-8852-D7D58BFA9213"><SST preparation.docx></span><span class="" style="font-family:Menlo-Regular; font-size:14px; font-style:normal; font-variant-caps:normal; font-weight:normal; letter-spacing:normal; text-align:start; text-indent:0px; text-transform:none; white-space:normal; word-spacing:0px; text-decoration:none; float:none; display:inline!important">_______________________________________________</span><br class="" style="font-family:Menlo-Regular; font-size:14px; font-style:normal; font-variant-caps:normal; font-weight:normal; letter-spacing:normal; text-align:start; text-indent:0px; text-transform:none; white-space:normal; word-spacing:0px; text-decoration:none">
<span class="" style="font-family:Menlo-Regular; font-size:14px; font-style:normal; font-variant-caps:normal; font-weight:normal; letter-spacing:normal; text-align:start; text-indent:0px; text-transform:none; white-space:normal; word-spacing:0px; text-decoration:none; float:none; display:inline!important">3dem
mailing list</span><br class="" style="font-family:Menlo-Regular; font-size:14px; font-style:normal; font-variant-caps:normal; font-weight:normal; letter-spacing:normal; text-align:start; text-indent:0px; text-transform:none; white-space:normal; word-spacing:0px; text-decoration:none">
<a href="mailto:3dem@ncmir.ucsd.edu" class="" style="font-family:Menlo-Regular; font-size:14px; font-style:normal; font-variant-caps:normal; font-weight:normal; letter-spacing:normal; orphans:auto; text-align:start; text-indent:0px; text-transform:none; white-space:normal; widows:auto; word-spacing:0px">3dem@ncmir.ucsd.edu</a><br class="" style="font-family:Menlo-Regular; font-size:14px; font-style:normal; font-variant-caps:normal; font-weight:normal; letter-spacing:normal; text-align:start; text-indent:0px; text-transform:none; white-space:normal; word-spacing:0px; text-decoration:none">
<a href="https://urldefense.proofpoint.com/v2/url?u=https-3A__eur03.safelinks.protection.outlook.com_-3Furl-3Dhttps-253A-252F-252Fmail.ncmir.ucsd.edu-252Fmailman-252Flistinfo-252F3dem-26data-3D04-257C01-257C-257Cd583a9cb213e43961e6908d994a5c5b2-257C4eed7807ebad415aa7a99170947f4eae-257C0-257C1-257C637704262000906937-257CUnknown-257CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0-253D-257C3000-26sdata-3DY2z31yPKx97rd-252BuMuq7rLGW8RaYm79J0D08tfzcK8Hk-253D-26reserved-3D0&d=DwMGaQ&c=-35OiAkTchMrZOngvJPOeA&r=L7-zyQ-04fFCMRqzLIOnx7H0exGZHwIQe_wMPuY600I&m=tI8xjaUudfF6hGCXLS5V5WtJs6dKaA3BCqrcotjccN8&s=W6fuHRA7Bv5psQfDalEGbfNLrzeVSZUjW4SjmqMP-2w&e=" originalsrc="https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem" shash="ewPINTb0OQbVl/aui79gNYDCJLVEd2zSgCQ0juuZ+gzM9AOS6+BAHnhkJZjrcOzXY9rl8iHMG7TGSTgbcnO8GC+zCRVcAmkrQ9oBJZE4zwYKDU8Mmc7kirqYIDfcjBJkmEfsTLDe5FKD4EM2ztowSEdXSGAsZ9MnPpr+7AykBD8=" class="" style="font-family:Menlo-Regular; font-size:14px; font-style:normal; font-variant-caps:normal; font-weight:normal; letter-spacing:normal; orphans:auto; text-align:start; text-indent:0px; text-transform:none; white-space:normal; widows:auto; word-spacing:0px">https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem</a></div>
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<p style="color:rgb(112,113,115);font-family: 'Trebuchet MS', 'Lucida Grande'; font-style: italic; font-size: 10pt;">
The Francis Crick Institute Limited is a registered charity in England and Wales no. 1140062 and a company registered in England and Wales no. 06885462, with its registered office at 1 Midland Road London NW1 1AT</p>
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