https://eln.excellgen.com/index.php?action=history&feed=atom&title=1612-pCold-7His-GST-%28E%29 1612-pCold-7His-GST-(E) - Revision history 2026-10-18T13:42:48Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1612-pCold-7His-GST-(E)&diff=8454&oldid=prev Paul.xu at 19:24, 5 March 2013 2013-03-05T19:24:05Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1612-pCold-7His-GST-(E)&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1612-pCold-7His-GST-(E) <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2013-1-22---2013-1-28 <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:1612&#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:1612 pcr1 1-24-2013.jpg]] <br /> <br /> PCR1:make promoter by gene synthesis <br /> <br /> Cold-1 Cold-2 Cold-3 Cold-4 Cold-5 Cold-6 Cold-7 Cold-8&lt;br&gt; <br /> <br /> pCold-R&lt;br&gt; <br /> <br /> [[Image:1608+1610+1611+1612+1613 pcr 1-10-2013.JPG]]&lt;br&gt; <br /> <br /> PCR2: PCR GST from pDB.His.GST&lt;br&gt;7His-GST-F&lt;br&gt;GST-Enterokinase-Nde1-R&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> [[Image:1612 pcr 1-25-2013.jpg]]&lt;br&gt; <br /> <br /> Join PCR products using &lt;br&gt;Cold-1 + GST-Enterokinase-Nde1-R&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> [[Image:1612 clone 1-25-2013.jpg]]&lt;br&gt; <br /> <br /> Clone into v19 pET28-cold using BglII + Nde1&lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2013-1-28 <br /> <br /> Make miniprep and digest with BglII + Nde1&lt;br&gt; <br /> <br /> [[Image:1471+1612 digest detect 1-28-2013.JPG]]&lt;br&gt; <br /> <br /> [[Image:1612 digest detect 1-28-2013.JPG]] <br /> <br /> &lt;br&gt; <br /> <br /> &amp;lt;br&amp;gt; <br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> *&amp;amp;amp;amp;amp;lt;br&amp;amp;amp;amp;amp;gt;<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> &amp;amp;amp;amp;amp;lt;br&amp;amp;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 /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Cloning of 1612-pCold-7His-GST-(E)&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1612-pCold-7His-GST-(E) <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2013-2-1----2013-2-4 <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:1612&#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&lt;br&gt; ===<br /> <br /> pcr1&lt;br&gt; <br /> <br /> PCR from v19 pET28-cold using <br /> <br /> Cold-BglII-F GGATCGAGATCTGCCGATTAATCATAAATATG&lt;br&gt;pCold-R atgatgatgatgatgatgatgcactttgtgattcatG&lt;br&gt; <br /> <br /> [[Image:1612+1614+1629 pcr1 2-1-2013.JPG]] <br /> <br /> &lt;br&gt; <br /> <br /> PCR2: PCR GST from pDB.His.GST&lt;br&gt;7His-GST-F&lt;br&gt;GST-Enterokinase-Nde1-R&lt;br&gt; <br /> <br /> [[Image:1608+1610+1611+1612+1613 pcr 1-10-2013.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> Join PCR products using &lt;br&gt;Cold-BglII-F + GST-Enterokinase-Nde1-R&lt;br&gt; <br /> <br /> [[Image:1612+1614+1629 pcr 2-1-2013.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2013-1-28 <br /> <br /> Make miniprep and digest with BglII + Nde1&lt;br&gt; <br /> <br /> [[Image:1612+1614+1629 digest detect 2-4-2013.JPG]]&lt;br&gt; <br /> <br /> 2013-3-5&lt;br&gt; <br /> <br /> lane 7: digest with Nhe1<br /> <br /> [[Image:1542+1662+1673_clone_3-5-2013.JPG]]<br /> <br /> &lt;br&gt; <br /> <br /> &amp;lt;br&amp;gt; <br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> *&amp;amp;amp;amp;amp;lt;br&amp;amp;amp;amp;amp;gt;<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> &amp;amp;amp;amp;amp;lt;br&amp;amp;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 /> [[Category:Cloning]]</div> Paul.xu