https://eln.excellgen.com/index.php?action=history&feed=atom&title=1629_pCold-GST-%283C%29 1629 pCold-GST-(3C) - Revision history 2026-10-18T11:23:23Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1629_pCold-GST-(3C)&diff=8111&oldid=prev Paul.xu at 17:49, 4 February 2013 2013-02-04T17:49:07Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;C onstruct&amp;nbsp; pCold-GST-(3C) vector&amp;nbsp;1629&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1629-pCold-GST-(3C) <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2013-1-25---2013-1- 26 <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:1629&#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 /> pcr1 <br /> <br /> Make cold promoter by gene synthesis&lt;br&gt; <br /> <br /> &lt;br&gt;&lt;br&gt; <br /> <br /> [[Image:1611pcr1+1427+1429pcr1 1-25-2013.jpg]] <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> pcr2&lt;br&gt;PCR GST from pDB.His.GST&lt;br&gt;Cold-GST-F+GST-3C-Nde1-R&lt;br&gt; <br /> <br /> [[Image:1625 1626 1629 PCR 2013 1 25.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> pcr1+pcr2 <br /> <br /> [[Image:1611+1614+1615+1629 pcr 1-28-2013.JPG]] <br /> <br /> &lt;br&gt; <br /> <br /> [[Image:1611+1614+1615+1629 clone 1-29-2013.JPG]]&lt;br&gt; <br /> <br /> digest pcr product&amp;nbsp; with BglII&amp;nbsp; + Nde1 <br /> <br /> &lt;br&gt;Clone into v19 pET28-cold using BglII + Nde1&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 /> == &#039;&#039;&#039;C onstruct&amp;nbsp; pCold-GST-(3C) vector&amp;nbsp;1629&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1629-pCold-GST-(3C) <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:1629&#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 /> pcr1 <br /> <br /> PCR from v19 pET28-cold using <br /> <br /> Cold-BglII-F + Cold-GST-R&lt;br&gt; <br /> <br /> [[Image:1612+1614+1629 pcr1 2-1-2013.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> pcr2&lt;br&gt;PCR GST from pDB.His.GST&lt;br&gt;Cold-GST-F+GST-3C-Nde1-R&lt;br&gt; <br /> <br /> [[Image:1625 1626 1629 PCR 2013 1 25.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> pcr1+pcr2 <br /> <br /> Cold-BglII-F+GST-3C-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: 2013-2-4 <br /> <br /> Make miniprep and digest with BglII + Nde1 &lt;br&gt; <br /> <br /> [[Image:1612+1614+1629_digest_detect_2-4-2013.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 /> [[Category:Cloning]]</div> Paul.xu