<html><head><meta http-equiv="Content-Type" content="text/html charset=us-ascii"></head><body style="word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space;">Hi John,<div><span class="Apple-tab-span" style="white-space:pre"> </span>The maximum volume depends on whether you are using the SedVel50 or SedVel60 version.You have to fill the sides so that the matched menisci (i.e. the sample and the reference menisci) are both above the capillary (the one that starts on the reference side, runs down the middle of the center rib and into the bottom of the sample side).</div><div>Otherwise, the reference buffer will stop flowing to the sample side when the meniscus gets to the capillary and too little buffer will have been transferred. And the menisci will not be matched.</div><div><br></div><div>You will have to experiment a little to find the sweet spot. I was using the SedVel60 version and - I may not be remembering correctly but I think I used 440 uL on the reference side and 420 uL on the sample side. So that when matched there was 430 uL on each side and my menisci were at 5.90 cm. This gives the longest sedimentation path, if you will. It's wasteful to use a smaller volume: the longer sedimentation path gives you more resolution. Each run is expensive, so you might as well get the most out of it.</div><div><br></div><div>Walter<br><div><div>On Dec 18, 2014, at 14:23, John Sumida <<a href="mailto:jpsumida@U.WASHINGTON.EDU">jpsumida@U.WASHINGTON.EDU</a>> wrote:</div><br class="Apple-interchange-newline"><blockquote type="cite"><meta http-equiv="Content-Type" content="text/html; charset=us-ascii"><meta name="Generator" content="Microsoft Word 14 (filtered medium)"><!--[if !mso]><style>v\:* {behavior:url(#default#VML);}
o\:* {behavior:url(#default#VML);}
w\:* {behavior:url(#default#VML);}
.shape {behavior:url(#default#VML);}
</style><![endif]--><style><!--
/* Font Definitions */
@font-face
{font-family:Calibri;
panose-1:2 15 5 2 2 2 4 3 2 4;}
@font-face
{font-family:Tahoma;
panose-1:2 11 6 4 3 5 4 4 2 4;}
/* Style Definitions */
p.MsoNormal, li.MsoNormal, div.MsoNormal
{margin:0in;
margin-bottom:.0001pt;
font-size:11.0pt;
font-family:"Calibri","sans-serif";}
a:link, span.MsoHyperlink
{mso-style-priority:99;
color:blue;
text-decoration:underline;}
a:visited, span.MsoHyperlinkFollowed
{mso-style-priority:99;
color:purple;
text-decoration:underline;}
p.MsoAcetate, li.MsoAcetate, div.MsoAcetate
{mso-style-priority:99;
mso-style-link:"Balloon Text Char";
margin:0in;
margin-bottom:.0001pt;
font-size:8.0pt;
font-family:"Tahoma","sans-serif";}
span.BalloonTextChar
{mso-style-name:"Balloon Text Char";
mso-style-priority:99;
mso-style-link:"Balloon Text";
font-family:"Tahoma","sans-serif";}
span.EmailStyle19
{mso-style-type:personal;
font-family:"Courier New";
color:windowtext;}
span.EmailStyle20
{mso-style-type:personal;
font-family:"Arial","sans-serif";
color:blue;
font-weight:normal;
font-style:normal;
text-decoration:none none;}
span.EmailStyle21
{mso-style-type:personal-reply;
font-family:"Courier New";
color:blue;}
.MsoChpDefault
{mso-style-type:export-only;
font-size:10.0pt;}
@page WordSection1
{size:8.5in 11.0in;
margin:1.0in 1.0in 1.0in 1.0in;}
div.WordSection1
{page:WordSection1;}
--></style><!--[if gte mso 9]><xml>
<o:shapedefaults v:ext="edit" spidmax="1026" />
</xml><![endif]--><!--[if gte mso 9]><xml>
<o:shapelayout v:ext="edit">
<o:idmap v:ext="edit" data="1" />
</o:shapelayout></xml><![endif]--><div lang="EN-US" link="blue" vlink="purple"><div class="WordSection1"><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New";color:blue">Dear RASMB<o:p></o:p></span></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New";color:blue"> </span></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New";color:blue">Thank you for your responses to my question. For the Stafford meniscus matching centerpieces what is the max volume that you can fill these centerpieces to?<o:p></o:p></span></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New";color:blue"> </span></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New";color:blue">Best regards,<o:p></o:p></span></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New";color:blue">John Sumida<o:p></o:p></span></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New";color:blue">University of Washington<o:p></o:p></span></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New";color:blue"> </span></p><div><div style="border:none;border-top:solid #B5C4DF 1.0pt;padding:3.0pt 0in 0in 0in"><p class="MsoNormal"><b><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif"">From:</span></b><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif""> RASMB [<a href="mailto:rasmb-bounces@list.rasmb.org">mailto:rasmb-bounces@list.rasmb.org</a>] <b>On Behalf Of </b>Karl Maluf<br><b>Sent:</b> Thursday, December 18, 2014 11:03 AM<br><b>To:</b> <a href="mailto:rasmb@rasmb.org">rasmb@rasmb.org</a><br><b>Subject:</b> Re: [RASMB] Centerpiece question<o:p></o:p></span></p></div></div><p class="MsoNormal"><o:p> </o:p></p><p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Arial","sans-serif";color:blue">The other issue with the meniscus matching centerpeices is to be careful to not overfill the buffer channel higher than the top capillary. This prevents equalization of the pressures between the two channels, and can inhibit the desired meniscus matching.</span><span style="font-size:12.0pt;font-family:"Times New Roman","serif""><o:p></o:p></span></p><div><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Times New Roman","serif""> <o:p></o:p></span></p></div><div><p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Arial","sans-serif";color:blue">Karl</span><span style="font-size:12.0pt;font-family:"Times New Roman","serif""><o:p></o:p></span></p></div><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Times New Roman","serif""> </span></p><div class="MsoNormal" align="center" style="text-align:center"><span style="font-size:12.0pt;font-family:"Times New Roman","serif""><hr size="2" width="100%" align="center"></span></div><p class="MsoNormal" style="margin-bottom:12.0pt"><b><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif"">From:</span></b><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif""> RASMB [<a href="mailto:rasmb-bounces@list.rasmb.org">mailto:rasmb-bounces@list.rasmb.org</a>] <b>On Behalf Of </b>John Philo<br><b>Sent:</b> Thursday, December 18, 2014 10:49 AM<br><b>To:</b> 'John Sumida'; <a href="mailto:rasmb@rasmb.org">rasmb@rasmb.org</a><br><b>Subject:</b> Re: [RASMB] Centerpiece question</span><span style="font-size:12.0pt;font-family:"Times New Roman","serif""><o:p></o:p></span></p><p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Arial","sans-serif";color:blue">John, in my view the advantages of the Stafford meniscus-matching design (available from Spin Analytical) over the synthetic boundary centerpieces are that the long capillary design inhibits diffusion between the two channels and any inadvertent fluid transfer during loading and handling. The drawback of that long channel though is that it is harder to get the transfer needed to match the heights (it takes more force)---I find I may need to go to 10-12 K rpm. For some of the virus samples we run that is >= to the run speed, so you have to do it very quickly to avoid significant sedimentation during the meniscus matching spin.<o:p></o:p></span></p><p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Arial","sans-serif";color:blue"> </span></p><p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Arial","sans-serif";color:blue">John<o:p></o:p></span></p><p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Arial","sans-serif";color:blue"> </span></p><div><div style="border:none;border-top:solid #E1E1E1 1.0pt;padding:3.0pt 0in 0in 0in"><p class="MsoNormal"><b>From:</b> RASMB [<a href="mailto:rasmb-bounces@list.rasmb.org">mailto:rasmb-bounces@list.rasmb.org</a>] <b>On Behalf Of </b>John Sumida<br><b>Sent:</b> Thursday, December 18, 2014 9:45 AM<br><b>To:</b> <a href="mailto:rasmb@rasmb.org">rasmb@rasmb.org</a><br><b>Subject:</b> [RASMB] Centerpiece question<o:p></o:p></p></div></div><p class="MsoNormal"><o:p> </o:p></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New"">Dear RASMB,<o:p></o:p></span></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New""> </span></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New"">For interference measurements where matching the sample and reference channel menisci is essential, is there a preference for either the meniscus matching centerpiece versus synthetic boundary centerpiece. We would be interested to hear about pros and cons.<o:p></o:p></span></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New""> </span></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New"">Thank you and happy holidays.<o:p></o:p></span></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New""> </span></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New"">Best regards,<o:p></o:p></span></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New""> </span></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New"">John Sumida, Ph.D.<o:p></o:p></span></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New""><a href="http://depts.washington.edu/cidb4bio/index.shtml"><span style="color:windowtext">Analytical Biopharmacy Core Facility</span></a><o:p></o:p></span></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New"">University of Washington<o:p></o:p></span></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New""><a href="http://www.moles.washington.edu/"><span style="color:windowtext">Molecular Engineering & Sciences Institute, G22</span></a><o:p></o:p></span></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New"">3946 West Stevens Way NE<o:p></o:p></span></p><p class="MsoNormal"><span style="font-size:12.0pt;font-family:"Courier New"">Seattle WA 98195-1653<o:p></o:p></span></p><p class="MsoNormal"><span style=""> </span></p></div></div>_______________________________________________<br>RASMB mailing list<br><a href="mailto:RASMB@list.rasmb.org">RASMB@list.rasmb.org</a><br>http://list.rasmb.org/listinfo.cgi/rasmb-rasmb.org<br></blockquote></div><br><div apple-content-edited="true"><br></div>
<br></div></body></html>