https://eln.excellgen.com/index.php?action=history&feed=atom&title=1626_pET28-cold-NRF2 1626 pET28-cold-NRF2 - Revision history 2026-10-18T12:08:04Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1626_pET28-cold-NRF2&diff=9298&oldid=prev Paul.xu at 19:40, 29 May 2013 2013-05-29T19:40:24Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1626 pET28-cold-NRF2&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1626 pET28-cold-NRF2 <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2013-1-25---2013-1-28&amp;nbsp; <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:1626&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039; yingying <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === PCR ===<br /> <br /> [[Image:1625 1626 1629 PCR 2013 1 25.JPG]]&lt;br&gt; <br /> <br /> 1/27/2013 <br /> <br /> [[Image:1625 1626 PCR digestion 1 27 2013.jpg]]&lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2013-1- <br /> <br /> Make miniprep and digest with &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 /> == &#039;&#039;&#039;Cloning of 1626 pET28-cold-NRF2&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1626 pET28-cold-NRF2 <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2013-1-29--2013-1-31 <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:1626&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039; yingying <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === PCR ===<br /> <br /> [[Image:1625+1626 pcr 1-29-2013.JPG]]&lt;br&gt; <br /> <br /> [[Image:1625+1626 clone 1-29-2013.JPG]]&lt;br&gt; <br /> <br /> clone into v19 pET28-cold vector using Nde1 + Xho1&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2013-1-31 <br /> <br /> Make miniprep and digest with Nde1 + Xho1&lt;br&gt; <br /> <br /> [[Image:1625+1626 digest detect 1-31-2013.JPG]] <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 /> == &#039;&#039;&#039;Protein Expression&#039;&#039;&#039; ==<br /> <br /> [[1625 pCold-GST-(3c)-KEAP1#Protein_Expression|1625_pCold-GST-(3c)-KEAP1#Protein_Expression]] <br /> <br /> == &#039;&#039;&#039;Cloning of 1626 pET28-cold-NRF2&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1626 pET28-cold-NRF2 <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2013-2-27--2013-3-1 <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:1626&#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:1626 PCR 2-27-2013.JPG]]&lt;br&gt; <br /> <br /> F2+R <br /> <br /> NRF2-Nde1-Nhe1-F2&lt;br&gt;catagc CATatg GCTAGC atggacttggagctgccgccgccg&lt;br&gt;NRF2-Kpn1-Xho1-R tgatcgCTCGAGGGTACCctagtttttcttaacatctggcttc&lt;br&gt; <br /> <br /> [[Image:1626 clone 2-27-2013.JPG]]&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: 2013-3-1 <br /> <br /> Make miniprep and digest with&amp;nbsp; Nde1 + Xho1 <br /> <br /> [[Image:1626 digest detect 3-1-2013.JPG]]&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 /> *&amp;amp;amp;lt;br&amp;amp;amp;gt;<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> <br /> <br /> <br /> <br /> == protein expression result of NRF2 clones &lt;br&gt; ==<br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot; width=&quot;475&quot;<br /> |-<br /> | clone ID&lt;br&gt; <br /> | expression&lt;br&gt; <br /> | gel reslut&lt;br&gt; <br /> | &lt;br&gt;<br /> |-<br /> | 1626, pET28-cold-NRF2&lt;br&gt; <br /> | no&lt;br&gt; <br /> | [[1625 1626 1656]]&lt;br&gt; <br /> | &lt;br&gt;<br /> |-<br /> | 1661,pT7A3-MBP-(3c)-NRF2&lt;br&gt; <br /> | yes,&lt;br&gt; <br /> | [[1661 1674 1683]]&lt;br&gt; <br /> | &lt;br&gt;<br /> |-<br /> | 1674,pCold-GST-(3c)-NRF2&lt;br&gt; <br /> | need to use GST beads&lt;br&gt; <br /> | [[1661 1674 1683]]&lt;br&gt; <br /> | &lt;br&gt;<br /> |-<br /> | 1683,pCold-MBP-(3C)-NRF2&lt;br&gt; <br /> | yes, &lt;br&gt; <br /> | [[1661 1674 1683]]&lt;br&gt; <br /> | &lt;br&gt;<br /> |-<br /> | 1698,pT7A3-NRF2&lt;br&gt; <br /> | no&lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt;<br /> |-<br /> | 1719,pT7A3-MBP-(E3C)-NRF2&lt;br&gt; <br /> | no&lt;br&gt; <br /> | [[626 1717 1719 1720]]&lt;br&gt; <br /> | &lt;br&gt;<br /> |-<br /> | 1748,pCold-8His-MBP-NRF2-6His&lt;br&gt; <br /> | only MBP&lt;br&gt; <br /> | [[1676 1726 1708 1748]]&lt;br&gt; <br /> | &lt;br&gt;<br /> |}<br /> <br /> &lt;br&gt; <br /> <br /> [[Category:Cloning]]</div> Paul.xu