https://eln.excellgen.com/index.php?action=history&feed=atom&title=1427-pLVX-Kozak-loxP-TurboRFP-ODC-loxP-TurboGFP 2026-10-18T05:19:47Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1427-pLVX-Kozak-loxP-TurboRFP-ODC-loxP-TurboGFP&diff=7127&oldid=prev 2012-11-20T15:48:57Z

<p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1427-pLVX-Kozak-loxP-TurboRFP-ODC-loxP-TurboGFP&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1427-pLVX-Kozak-loxP-TurboRFP-ODC-loxP-TurboGFP <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2012-11-8---2012-11-14 <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:1427&#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:1427 PCR 11-8-2012.JPG]]&lt;br&gt; <br /> <br /> PCR using 81 &#039;&#039;&#039;TurboGFP&#039;&#039;&#039; as template, and &lt;br&gt;the new TurboGFP-Xho1-F (without Kozak seqeunce and start codon)&lt;br&gt;and&lt;br&gt;TurboGFP-Xba1-R&lt;br&gt; <br /> <br /> [[Image:1427 clone 11-9-2012.JPG]]&lt;br&gt; <br /> <br /> {| width=&quot;385&quot; cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot;<br /> |-<br /> | lane&lt;br&gt; <br /> | 1&lt;br&gt; <br /> | 2&lt;br&gt;<br /> |-<br /> | &lt;br&gt; <br /> | PCR product&lt;br&gt; <br /> | 1425-pLVX-Kozak-loxP-TurboRFP-ODC-loxP-Gluc&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;&amp;nbsp;&amp;nbsp;&amp;nbsp; Xho1+ Xba1&lt;br&gt;<br /> |}<br /> <br /> &lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2012-11-13 <br /> <br /> Make miniprep and digest with&amp;nbsp; Xho1+ Xba1 <br /> <br /> [[Image:1427 digest detect 11-13-2012.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> 2012-11-14&lt;br&gt; <br /> <br /> 6 clonies PCR screening:new TurboGFP-Xho1-F+PGK-P-R-Seq&lt;br&gt; <br /> <br /> [[Image:1427 PCR Screening 11-14-2012.JPG]] <br /> <br /> clone 3,4,5,6 may be right ,need to sequence <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;amp;lt;br&amp;amp;amp;amp;gt;<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> &amp;amp;amp;amp;lt;br&amp;amp;amp;amp;gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot;<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 /> {|<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> == &#039;&#039;&#039;Cloning of 1427-pLVX-Kozak-loxP-TurboRFP-ODC-loxP-TurboGFP&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1427-pLVX-Kozak-loxP-TurboRFP-ODC-loxP-TurboGFP <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2012-11-15---2012-11-18 <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:1427&#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:1425_PCR_11-15-2012.JPG]]&lt;br&gt;<br /> <br /> PCR &#039;&#039;&#039;loxP-TurboRFP-ODC-loxP &#039;&#039;&#039;from 1271 pLVX-loxP-TurboRFP-ODC-loxP-Gluc&lt;br&gt;<br /> <br /> LoxP-TurboRFP-BamH1-F1+LoxP-TurboRFP-BamH1-F2+LoxP-hGHpA-Xho1-R2&lt;br&gt; <br /> <br /> <br /> <br /> [[Image:567+1427 clone 11-15-2012.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> {| width=&quot;385&quot; cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot;<br /> |-<br /> | lane&lt;br&gt; <br /> | 1&lt;br&gt; <br /> | 2&lt;br&gt;<br /> |-<br /> | &lt;br&gt; <br /> | PCR product&lt;br&gt; <br /> | 1427-pLVX-Kozak-loxP-TurboRFP-ODC-loxP-TurboGFP&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;&amp;nbsp;&amp;nbsp;&amp;nbsp; BamH1 + Xho1&lt;br&gt;<br /> |}<br /> <br /> &lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2012-11-19 <br /> <br /> Make miniprep and digest with&amp;nbsp; BamH1 + Xho1 <br /> <br /> [[Image:1424+1427 digest detect 11-19-2012.JPG]]&lt;br&gt; <br /> <br /> Clone 1,2,3,4,5,6,7 need to sequencing <br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> *&amp;amp;amp;amp;lt;br&amp;amp;amp;amp;gt;<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> &amp;amp;amp;amp;lt;br&amp;amp;amp;amp;gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot;<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 /> {|<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> [[Category:Cloning]]</div>

Paul.xu