https://eln.excellgen.com/index.php?action=history&feed=atom&title=1424-pCMV5-IgK-ATR-Fc-WPRE-oriP 1424-pCMV5-IgK-ATR-Fc-WPRE-oriP - Revision history 2026-10-18T14:43:47Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1424-pCMV5-IgK-ATR-Fc-WPRE-oriP&diff=7322&oldid=prev Paul.xu at 14:47, 4 December 2012 2012-12-04T14:47:08Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1424-pCMV5-IgK-ATR-Fc-WPRE-oriP&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1424-pCMV5-IgK-ATR-Fc-WPRE-oriP <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2012-10-24---2012-10-26 <br /> <br /> &#039;&#039;&#039;Notebook:0&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID:1424&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039; yingying <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === cloning ===<br /> <br /> [[Image:1424 PCR 10-25-2012.JPG]]&lt;br&gt; <br /> <br /> not the right size <br /> <br /> &lt;br&gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot;<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> === Colony Screening ===<br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> *&amp;amp;amp;lt;br&amp;amp;amp;gt;<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> &amp;amp;amp;lt;br&amp;amp;amp;gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot;<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> &lt;br&gt; <br /> <br /> {|<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> &lt;br&gt; <br /> <br /> {|<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> &lt;br&gt; <br /> <br /> = &#039;&#039;&#039;Cloning of 1424-pCMV5-IgK-ATR-Fc-WPRE-oriP&#039;&#039;&#039; = =<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1424-pCMV5-IgK-ATR-Fc-WPRE-oriP <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2012-11-15---2012-11-19 <br /> <br /> &#039;&#039;&#039;Notebook:0&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID:1424&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039; yingying <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === cloning ===<br /> <br /> PCR using the following oligos: <br /> <br /> 1 ul of 10 uM: ATR-IgG-k-BamH1-F1 &lt;br&gt;1 ul of 1 uM: ATR-IgG-k-BamH1-F2 &lt;br&gt;1 ul of 1 uM: ATR-IgG-k-BamH1-F3 &lt;br&gt;1 ul of 1 uM: ATR-IgG-k-BamH1-F4 &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> 1 ul of 10 uM: ATR-Not1-R&lt;br&gt;&lt;br&gt; <br /> <br /> template 1456,pLP-CMV-ATR&lt;br&gt;&lt;br&gt; <br /> <br /> [[Image:ATR PCR 11-16-2012.JPG]][[Image:ATR PCR2 11-16-2012.JPG]][[Image:1456 plasmid QC 11-16-2012.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> [[Image:1424 clone 11-16-2012.JPG]]&lt;br&gt; <br /> <br /> {| width=&quot;399&quot; cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot;<br /> |-<br /> | lane&lt;br&gt; <br /> | 1&lt;br&gt; <br /> | 2&lt;br&gt;<br /> |-<br /> | &lt;br&gt; <br /> | PCR product&lt;br&gt; <br /> | 1387,pCMV5-hIgG-Hc-Dest-Fc(3c)-WPRE-OriP&lt;br&gt;<br /> |-<br /> | enzymes&lt;br&gt; <br /> | colspan=&quot;2&quot; | &amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; BamH1 + Not1&lt;br&gt;<br /> |}<br /> <br /> &lt;br&gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot;<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> === Colony Screening ===<br /> <br /> Date: 2012-11-19 <br /> <br /> Make miniprep and digest with BamH1 + Not1&lt;br&gt; <br /> <br /> [[Image:1424+1427 digest detect 11-19-2012.JPG]] <br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> *&amp;amp;amp;lt;br&amp;amp;amp;gt;<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> &amp;amp;amp;lt;br&amp;amp;amp;gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot;<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> &amp;amp;amp;lt;br&amp;amp;amp;gt; <br /> <br /> {| cellspacing=&quot;1&quot;<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> &amp;amp;lt;br&amp;amp;gt; <br /> <br /> {|<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> &amp;lt;br&amp;gt; <br /> <br /> {|<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> &lt;br&gt; <br /> <br /> {|<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> == &#039;&#039;&#039;Cloning of 1424-pCMV5-IgK-ATR-Fc-WPRE-oriP&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1424-pCMV5-IgK-ATR-Fc-WPRE-oriP <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2012-11-26 <br /> <br /> &#039;&#039;&#039;Notebook:0&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID:1424&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039; yingying <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === cloning ===<br /> <br /> PCR using the following oligos: <br /> <br /> 1 ul of 10 uM: ATR-IgG-k-BamH1-F1&lt;br&gt;1 ul of 1 uM: ATR-IgG-k-BamH1-F2&lt;br&gt;1 ul of 1 uM: ATR-IgG-k-BamH1-F3&lt;br&gt;1 ul of 1 uM: ATR-IgG-k-BamH1-F4 <br /> <br /> 1 ul of 10 uM: ATR-Not1-R <br /> <br /> template 1456,pLP-CMV-ATR&lt;br&gt; <br /> <br /> [[Image:1424 PCR 11-26-2012.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> [[Image:1424+1426+1472+1473+1474+1475+1476 clone 11-26-2012.JPG]]&lt;br&gt; <br /> <br /> after digestion, the size became small <br /> <br /> {| width=&quot;414&quot; cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot;<br /> |-<br /> | &lt;br&gt; <br /> | lane 1&lt;br&gt; <br /> | lane 2&lt;br&gt;<br /> |-<br /> | &lt;br&gt; <br /> | PCR product&lt;br&gt; <br /> | 1391 pCMV5-TPA-TagGFP-(3c)-Fc-WPRE-OriP&lt;br&gt;<br /> |-<br /> | enzymes &lt;br&gt; <br /> | colspan=&quot;2&quot; | BamH1 + Not1&lt;br&gt;<br /> |}<br /> <br /> &lt;br&gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot;<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> === Colony Screening ===<br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> *&amp;amp;amp;lt;br&amp;amp;amp;gt;<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> &amp;amp;amp;lt;br&amp;amp;amp;gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot;<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> &lt;br&gt; <br /> <br /> {|<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> &lt;br&gt; <br /> <br /> {|<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> &lt;br&gt; <br /> <br /> <br /> <br /> [[Category:Cloning]]</div> Paul.xu