https://eln.excellgen.com/index.php?action=history&feed=atom&title=1265_pEC-Tks 1265 pEC-Tks - Revision history 2026-10-18T08:48:53Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1265_pEC-Tks&diff=8972&oldid=prev Paul.xu at 19:38, 25 April 2013 2013-04-25T19:38:32Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1265 pEC-Tks-&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of1265 pEC-Tks <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2013-3- <br /> <br /> &#039;&#039;&#039;Notebook:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID:1265&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039; Shanyi <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039;&lt;br&gt; ==<br /> <br /> === cloning ===<br /> <br /> Date:2013-3-&lt;br&gt; <br /> <br /> Gene synthesis 4 PCR fragments <br /> <br /> [[Image:1265firstpcr 03 26 2013.JPG]]&lt;br&gt; <br /> <br /> &amp;nbsp;then use Fragment1,2,3 ,4as template Tks-1-1+Tks-4-18 as primers <br /> <br /> [[Image:1265 1592pcr 03 27 2013.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; digest&amp;nbsp;: <br /> <br /> [[Image:DIGEST FOR 1265 1535.JPG]]&lt;br&gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot; style=&quot;width: 298px; height: 82px;&quot;<br /> |-<br /> | &lt;br&gt; <br /> | band 1,2&lt;br&gt; <br /> | band 3&lt;br&gt;<br /> |-<br /> | &lt;br&gt; <br /> | &amp;nbsp; PCR product&lt;br&gt; <br /> | 1454&lt;br&gt;<br /> |-<br /> | enzymes&lt;br&gt; <br /> | colspan=&quot;2&quot; | &amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; Nde1 + Kpn1&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: 2013 <br /> <br /> DIGEST WITH HindIIIshould get three bands <br /> <br /> [[Image:1535 1265digest.JPG]]&lt;br&gt; <br /> <br /> SEND 1265-2 TO SEQUENCE,missing one G&lt;br&gt; &lt;br&gt; &lt;br&gt; <br /> <br /> === cloning ===<br /> <br /> Date:2013-3-&lt;br&gt; <br /> <br /> PCR using 1265-2as template&lt;br&gt; <br /> <br /> use Tks-1-1 and Tks-2-10, Tks-2-11 and Tks-4-18 as primers <br /> <br /> [[Image:1265FIX firstPCR 04 17 2013.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> &amp;nbsp;then use Fragment1,2as template Tks-1-1+Tks-4-18 as primers <br /> <br /> [[Image:1265FIX PCR 04 22 2013.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; digest&amp;nbsp;: <br /> <br /> [[Image:DIGEST FOR1746 1265FIX 04 23 2013.JPG]] <br /> <br /> &lt;br&gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot; style=&quot;width: 298px; height: 82px;&quot;<br /> |-<br /> | &lt;br&gt; <br /> | band 5,6&lt;br&gt; <br /> | band 4&lt;br&gt;<br /> |-<br /> | &lt;br&gt; <br /> | &amp;nbsp; PCR product&lt;br&gt; <br /> | 1454&lt;br&gt;<br /> |-<br /> | enzymes&lt;br&gt; <br /> | colspan=&quot;2&quot; | &amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; Nde1 + Kpn1&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: 2013 <br /> <br /> DIGEST WITH HindIIIshould get three bands <br /> <br /> [[Image:1265fix_digest_04_25_2013.JPG‎]]&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 /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> [[Category:Cloning]]</div> Paul.xu