https://eln.excellgen.com/index.php?action=history&feed=atom&title=1150_pHis.MBP.TEV 1150 pHis.MBP.TEV - Revision history 2026-10-18T10:35:27Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1150_pHis.MBP.TEV&diff=4180&oldid=prev Paul.xu at 20:17, 12 May 2012 2012-05-12T20:17:22Z <p></p> <p><b>New page</b></p><div>Cloning of pHis.MBP.TEV(1150)&lt;br&gt;Purpose: to clone pHis.MBP.TEV&lt;br&gt;Date: 2012-5-1 to 2012-5-&lt;br&gt;Notebook: Xiaojing Wang&lt;br&gt;Page: 004237 <br /> <br /> Clone ID: 1150&lt;br&gt;Experimenters: Xiaojing Wang <br /> <br /> Results &lt;br&gt; <br /> <br /> Digest pHis.MBP 1116 with Nde1 + EcoR1 --&amp;gt; insert&lt;br&gt;Digest pHis-GST-TEV 1030 with Nde1 + EcoR1 --&amp;gt; vector (discard 660bp GST fragment ) <br /> <br /> [[Image:1150 i v digest 5.1.2012.JPG]]&lt;br&gt; <br /> <br /> Ligate and transform TOP10&lt;br&gt;Select 4 clones to mini-prep&lt;br&gt;1150 plasmids are digested by Nde1 + EcoR1&lt;br&gt; <br /> <br /> [[Image:1150digest 5.3.2012.JPG]]&lt;br&gt; <br /> <br /> No plasmid is correct.&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> repeat 5/4/2012&lt;br&gt; <br /> <br /> [[Image:1150 i v digest 5.4.2012.JPG]]&lt;br&gt; <br /> <br /> Ligate and transform TOP10&lt;br&gt;Select 6 clones to mini-prep&lt;br&gt;1150 plasmids are digested by Nde1 + EcoR1&lt;br&gt; <br /> <br /> [[Image:1150digest 5.7.2012.JPG]]&lt;br&gt; <br /> <br /> No plasmid is correct.&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> New method 5/9/2012&lt;br&gt;digest pHis.MBP 1116 with EcoR1 + Xho1 --&amp;gt;vector&lt;br&gt;Digest pHis-GST-TEV 1030 with EcoR1+ Xho1--&amp;gt; insert&lt;br&gt; <br /> <br /> [[Image:1150 i v digest 5.9.2012.JPG]]&lt;br&gt; <br /> <br /> --[[User:Paul.xu|Paul.xu]] 15:15, 12 May 2012 (EST) <br /> <br /> 5/12/2012. <br /> <br /> pick 5 colonies, make miniprep, digest with Nde1 or EcoR1 + Xho1. DNA marker is Fermentas 1kb plus (20000, 10000, 7000, &#039;&#039;&#039;5000&#039;&#039;&#039;, 4000, 3000, 2000, &#039;&#039;&#039;1500&#039;&#039;&#039;, 1000, 700, &#039;&#039;&#039;500,&#039;&#039;&#039; 400, 300, 200, 75).<br /> <br /> [[Image:1150 Nde1 5 12 2012.jpg]]&lt;br&gt; <br /> <br /> [[Image:1150 EcoR1 Xho1 5 12 2012.jpg]] --[[User:Paul.xu|Paul.xu]] 15:15, 12 May 2012 (EST) <br /> <br /> [[Category:Cloning]]</div> Paul.xu