https://eln.excellgen.com/index.php?action=history&feed=atom&title=2315_pEC2-TEV 2315 pEC2-TEV - Revision history 2026-10-18T23:05:46Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=2315_pEC2-TEV&diff=13061&oldid=prev Excellgen: Created page with "== '''Cloning of 2315 pEC2-TEV''' == '''Purpose:''' Cloning of2315 pEC2-TEV'''==''' '''Date: '''2014-9---- '''Notebook:0''' '''Page:''' '''Clone ID:2315''' '''Experim..." 2014-09-11T16:57:25Z <p>Created page with &quot;== &#039;&#039;&#039;Cloning of 2315 pEC2-TEV&#039;&#039;&#039; == &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of2315 pEC2-TEV&#039;&#039;&#039;==&#039;&#039;&#039; &#039;&#039;&#039;Date: &#039;&#039;&#039;2014-9---- &#039;&#039;&#039;Notebook:0&#039;&#039;&#039; &#039;&#039;&#039;Page:&#039;&#039;&#039; &#039;&#039;&#039;Clone ID:2315&#039;&#039;&#039; &#039;&#039;&#039;Experim...&quot;</p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 2315 pEC2-TEV&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of2315 pEC2-TEV&#039;&#039;&#039;==&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2014-9---- <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:2315&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039;&amp;nbsp; shanyi <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === CLONING ===<br /> <br /> <br /> <br /> use the following oligos and 432 pMBP-7xHis-TEV (pMHTDelta238) as template<br /> <br /> <br /> <br /> &lt;br&gt;LysN-Tev-2&lt;br&gt;Sequence ATCGCCTCTAGATCCCTATCAGTGATAGAGATTG<br /> <br /> LysN-Tev-3&lt;br&gt;Sequence TGTGCTCAGTATCTCTATCACTGATAGGGATGTCAATCTCTATCACTGATAGGGATCT&lt;br&gt; (AGATCCCTATCAGTGATAGAGATTGACATCCCTATCAGTGATAGAGATACTGAGCACA)&lt;br&gt;LysN-Tev-4&lt;br&gt;Sequence CAGTGATAGAGATACTGAGCACATCAGCAGGACGCACTGACCGAATTGATTTAAGGAG<br /> <br /> LysN-Tev-5&lt;br&gt;Sequence CTTCCAGCCCGGCACCCATGGATCTTACCTCCTTAAATCAATTCGGTCAGT&lt;br&gt; (ACTGACCGAATTGATTTAAGGAGGTAAGATCCATGGGTGCCGGGCTGGAAG)<br /> <br /> &lt;br&gt;1 ul of 10 uM of &lt;br&gt;PlateName 2012_3_27&lt;br&gt;Pos D08&lt;br&gt;SequenceName LysN-Tev-seq-F&lt;br&gt;Sequence GTCGATGGATCCCTATTCATTCATGAGTTGAGTC&lt;br&gt;Year 2012&lt;br&gt;Date 3/30/2012<br /> <br /> gtcgatggatccCTATTCATTCATGAGTTGAGTC&lt;br&gt;(GACTCAACTCATGAATGAATAGggatcc atcgac)<br /> <br /> &lt;br&gt;LysN-Tev-2--&amp;gt;&amp;gt; TCTAGATCCCTATCAGTGATAGAGATTG CAGTGATAGAGATACTGAGCACATCAGCAGGACGCACTGACCGAATTGATTTAAGGAG LysN-Tev-4--&amp;gt;&amp;gt;&lt;br&gt;promoter TCTAGATCCCTATCAGTGATAGAGATTGACATCCCTATCAGTGATAGAGATACTGAGCACATCAGCAGGACGCACTGACCGAATTGATTTAAGGAGGTAAgATCC TEV--&amp;gt; ATGGGTGCCGGGCTGGAAG&lt;br&gt; (AGATCCCTATCAGTGATAGAGATTGACATCCCTATCAGTGATAGAGATACTGAGCACA) &amp;lt;&amp;lt;--LysN-Tev-3&lt;br&gt; ACTGACCGAATTGATTTAAGGAGGTAAGATCC ATGGGTGCCGGGCTGGAAG &amp;lt;&amp;lt;--LysN-Tev-5&lt;br&gt;<br /> <br /> &lt;br&gt;<br /> <br /> [[Image:2315_2166_2256PCR.JPG]]&lt;br&gt;<br /> <br /> <br /> <br /> BamH1 + Xba1&lt;br&gt;<br /> <br /> [[Image:DIGEST FOR 2315 2166 2256 2320 2332 2324.JPG]]&lt;br&gt;&lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2014_9&lt;br&gt; <br /> <br /> Make miniprep, use BamH1 + Xba1 to find positive clone&lt;br&gt; <br /> <br /> [[Image:2315DIGEST.JPG]]<br /> <br /> &lt;br&gt; <br /> <br /> digest with NdeI+Hindiii to verify <br /> <br /> [[Image:2135 2332 2324digest.JPG]] <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> *&lt;br&gt;<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> [[Category:Cloning]]</div> Excellgen