https://eln.excellgen.com/index.php?action=history&feed=atom&title=2227_pFastBac-Dual-VSVG-HIVGag-FLP-CD63-Flp 2227 pFastBac-Dual-VSVG-HIVGag-FLP-CD63-Flp - Revision history 2026-10-18T14:12:55Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=2227_pFastBac-Dual-VSVG-HIVGag-FLP-CD63-Flp&diff=11995&oldid=prev Paul.xu: Created page with "== '''Cloning of 2227 pFastBac-Dual-VSVG-HIVGag-FLP-CD63-Flp''' == '''Purpose:''' Cloning of 2227 pFastBac-Dual-VSVG-HIVGag-FLP-CD63-Flp '''Date: '''2014-6 '''Notebook:0'''..." 2014-06-12T19:23:33Z <p>Created page with &quot;== &#039;&#039;&#039;Cloning of 2227 pFastBac-Dual-VSVG-HIVGag-FLP-CD63-Flp&#039;&#039;&#039; == &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 2227 pFastBac-Dual-VSVG-HIVGag-FLP-CD63-Flp &#039;&#039;&#039;Date: &#039;&#039;&#039;2014-6 &#039;&#039;&#039;Notebook:0&#039;&#039;&#039;...&quot;</p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 2227 pFastBac-Dual-VSVG-HIVGag-FLP-CD63-Flp&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 2227 pFastBac-Dual-VSVG-HIVGag-FLP-CD63-Flp <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2014-6 <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:2227&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039; shanyi <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === cloning&lt;br&gt; ===<br /> <br /> 1. PCR CD63-Flpe from 2137 pCMV5-CD63-Flpe using &lt;br&gt;CD63-Sal1-F AAGACTGTCGACATGGCGGTGGAAGGAGGAATG&lt;br&gt;Flpe-Xba1-R GCAGGCTCTAGACTAGATCCGCCGGTTGATGTAG <br /> <br /> [[Image:2227 2228 2225 2220pcr2.JPG]] <br /> <br /> 2. PCR p10 promoter from pFastbac-Dual <br /> <br /> p10-Not1-F gctac GCGGCCGC GAGTATACGGACCTTTAATTC&lt;br&gt;p10-Sal1-CD63-R&lt;br&gt;(CATTTTATTTACAATCACTCGACGAAGACTGTCGAC)&lt;br&gt;GTCGACAGTCTTCGTCGAG <br /> <br /> TGATTGTAAATAAAATG&lt;br&gt; <br /> <br /> [[Image:2227 2228 2224 2225pcr1.JPG]] <br /> <br /> &lt;br&gt; <br /> <br /> 3. Join gel 2 fragments (maybe it is NOT necessary to purify two fragment on gel) using <br /> <br /> p10-Not1-F + Flpe-Xba1-R <br /> <br /> [[Image:2227 2228 2225 2220pcr3.JPG]]&lt;br&gt;4. clone into Not1 + Xba1 digested 2226 pFastBac-Dual-VSVG-HIVGag-FLP <br /> <br /> [[Image:Digest for 2227 22282nd.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2014-6 <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2014-6 <br /> <br /> mini prep and digest with EcoR1 <br /> <br /> [[Image:2112 2227 2228digest.JPG]] <br /> <br /> &lt;br&gt; verify using Not1 + Xba1 and BamHI&lt;br&gt; <br /> <br /> [[Image:1588REAL 2112 2227 2228DIGEST.JPG]] <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; &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 /> &amp;amp;amp;lt;br&amp;amp;amp;gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot;<br /> |-<br /> | &lt;br&gt;<br /> |}<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