https://eln.excellgen.com/index.php?action=history&feed=atom&title=1191_pCDNA3.1-Dest2 2026-10-18T02:24:39Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1191_pCDNA3.1-Dest2&diff=4974&oldid=prev 2012-07-06T12:54:32Z

<p></p> <p><b>New page</b></p><div>5/22/2012&lt;br&gt; <br /> <br /> digest pCDNA3.1-Dest 1190 with Not1 + BamH1 (Lane #3).&lt;br&gt;cut out vector (not the small fragment), purify with column, elute with 20 ul water. &lt;br&gt;Blunt end:&lt;br&gt;Add 2.5 ul buffer 4, 1 ul of 10 mM dNTPs. &lt;br&gt;Add 1 unit (0.3 ul of 3 u/ul) T4 DNA Polymerase&lt;br&gt;incubate 15 minutes at 12°C. &lt;br&gt;Purify:&lt;br&gt;Add gel extraction buffer and purify with column. &lt;br&gt;ligate and transform ccdb survival.&lt;br&gt; <br /> <br /> [[Image:1190 diges 5-22-2012.JPG]]&lt;br&gt; <br /> <br /> 5/24/2012 <br /> <br /> [[Image:1191 digest 5.24.2012.JPG]] <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Cloning of 1191 pCDNA3.1-Dest2&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of1191 pCDNA3.1-Dest2 <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2012-6-28---2012-7-5 <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:1191&#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:1235+1238_digest_6-26-2012.JPG]]&lt;br&gt; <br /> <br /> ligate to annealed oligos&lt;br&gt;GGCCGA ACGCCGATATTGAAGGTATT c&lt;br&gt;ct TGCGGCTATAACTTCCATAA gCTAG<br /> <br /> PfuMBP-seq-F&lt;br&gt;GGCCGA ACGCCGATATTGAAGGTATT c&lt;br&gt;GW-fix-B&lt;br&gt;GATCgAATACCTTCAATATCGGCGTtc&lt;br&gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot; style=&quot;width: 281px; height: 66px;&quot;<br /> |-<br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | band 4&lt;br&gt;<br /> |-<br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | 1238pCDNA3.1-Dest 1238&lt;br&gt;<br /> |-<br /> | enzymes&lt;br&gt; <br /> | colspan=&quot;2&quot; | &amp;nbsp;&amp;nbsp;&amp;nbsp;Not1 + BamH1&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; .&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: 2012-7-5<br /> <br /> Make miniprep and digest with Kpn1+Xho1&lt;br&gt; <br /> <br /> [[Image:1191_digest_detect_7-5-2012.JPG]]<br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> *&amp;amp;lt;br&amp;amp;gt;<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> <br /> [[Category:Cloning]]</div>

Paul.xu