https://eln.excellgen.com/index.php?action=history&feed=atom&title=Plasmid_MaxiPrep_by_Q_Sepharose 2026-10-19T00:24:27Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=Plasmid_MaxiPrep_by_Q_Sepharose&diff=6565&oldid=prev 2012-10-16T15:53:42Z

<p></p> <p><b>New page</b></p><div>Most economical, but need to optimize to remove endotoxin.<br /> <br /> <br /> <br /> Column: 20 ml of Q-Sepharose for 300 ml bacterial culture&lt;br&gt; <br /> <br /> === Wash Buffer: &lt;br&gt; ===<br /> <br /> 1. 40% isopropanol and 1.5 M NaCl&lt;br&gt; <br /> <br /> 2.&amp;nbsp;30% isopropanol&amp;nbsp; 1M NaCl &lt;br&gt; <br /> <br /> 3. 45% isopropanol 0.75M&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> Qiagen Q column: <br /> <br /> RNA will be eluted from column at 1.0 M NaCl at pH7. DNA will be eluted at 1.4 ~ 1.6 M NaCl. pH is very critical. <br /> <br /> === Buffer QC&lt;br&gt; ===<br /> <br /> 1.0 M NaCl&lt;br&gt;50 mM MOPS&lt;br&gt;Adjust pH to 7.0 with HCl&lt;br&gt;15% (v/v) isopropanol&lt;br&gt;Store indefinitely at room temperature&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> === Buffer QF&lt;br&gt; ===<br /> <br /> &lt;br&gt;1.25 M NaCl&lt;br&gt;50 mM Tris⋅Cl&lt;br&gt;15% isopropanol&lt;br&gt;Adjust pH to 8.5 with HCl&lt;br&gt;Store indefinitely at room temperature&lt;br&gt;&lt;br&gt; <br /> <br /> === Buffer QBT ===<br /> <br /> &lt;br&gt;750 mM NaCl&lt;br&gt;50 mM 3-(N-morpholino)propanesulfonic acid (MOPS)&lt;br&gt;Adjust pH to 7.0 with HCl&lt;br&gt;15% (v/v) isopropanol&lt;br&gt;0.15% (v/v) Triton X-100&lt;br&gt;Store indefinitely at room temperature&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> == Endotoxin Removal ==<br /> <br /> Dilute Eluted DNA 1:1 with 4M ammonium sulfate buffer then loaded onto a 2.6×2.5 cm Octyl HIC column equilibrated in 2M ammonium sulfate buffer. <br /> <br /> &lt;br&gt; <br /> <br /> [[Category:Protocols]]</div>

Paul.xu