https://eln.excellgen.com/index.php?action=history&feed=atom&title=306_pAAV-EF1-MCS 306 pAAV-EF1-MCS - Revision history 2026-10-18T17:31:44Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=306_pAAV-EF1-MCS&diff=1477&oldid=prev Hylei: Created page with "== '''Cloning of EF to pAAV-CMV-MCS''' == '''Purpose:''' Cloning of EF to pAAV-CMV-MCS. '''Date: '''2011-8-8 '''Notebook:''' '''Page:''' '''Clone ID:''' '''Experimenter..." 2011-08-08T20:36:27Z <p>Created page with &quot;== &#039;&#039;&#039;Cloning of EF to pAAV-CMV-MCS&#039;&#039;&#039; == &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of EF to pAAV-CMV-MCS. &#039;&#039;&#039;Date: &#039;&#039;&#039;2011-8-8 &#039;&#039;&#039;Notebook:&#039;&#039;&#039; &#039;&#039;&#039;Page:&#039;&#039;&#039; &#039;&#039;&#039;Clone ID:&#039;&#039;&#039; &#039;&#039;&#039;Experimenter...&quot;</p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of EF to pAAV-CMV-MCS&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of EF to pAAV-CMV-MCS. <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2011-8-8 <br /> <br /> &#039;&#039;&#039;Notebook:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039;<br /> <br /> &#039;&#039;&#039;Clone ID:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039; Haiyan <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === Subcloning ===<br /> <br /> Date: 2011-6-8 <br /> <br /> <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2011-8-8&lt;br&gt; <br /> <br /> Make miniprep and digest with Mlui+ecori&lt;br&gt; <br /> <br /> [[Image:0808_295_306_0808.JPG]] &lt;br&gt; <br /> <br /> Lane 1: marker &lt;br&gt; <br /> <br /> Lane 2-5: 295_pAAV-Syn-MCS, Mlui+ecori&lt;br&gt; <br /> <br /> Lane 6-8: 306_pAAV-EF-MCS, Mlui+ecori&lt;br&gt; <br /> <br /> Looks 295 no positive clone, 306 looks have insert, need to do pcr additional confirm.&lt;br&gt;<br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> *PKAalpha 3+4 has one internal Xho1 site. GFP-Puro can be cut out with Xho1 + Mlu1. So Lane 1 has correct number of bands. <br /> *PKA alpha 3+4 with CMV promoter can be cut out with Nde1+ BamH1. Lane 5 has correct number of bands. <br /> *lane 6 maybe good (incomplete digestion) <br /> *Sequencing Result:&lt;br&gt;<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> Restriction digestion &lt;br&gt; <br /> <br /> shows&amp;nbsp; pLentiGP clone #1 may be good, &lt;br&gt; <br /> <br /> pLVGP-PKAalpha 3+4 is good&lt;br&gt; <br /> <br /> === ===<br /> <br /> [[User:Paul.xu|Paul.xu]] 08:23, 15 June 2011 (EST)&lt;br&gt; <br /> <br /> [[Category:Cloning]]</div> Hylei