https://eln.excellgen.com/index.php?action=history&feed=atom&title=363%2B1336_gateaway_transfection_on_9-14-2012 363+1336 gateaway transfection on 9-14-2012 - Revision history 2026-10-18T03:32:39Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=363%2B1336_gateaway_transfection_on_9-14-2012&diff=6190&oldid=prev Paul.xu: moved 1363+1336 gateaway transfection on 9-14-2012 to 363+1336 gateaway transfection on 9-14-2012 over redirect 2012-09-23T20:46:14Z <p>moved <a href="/index.php?title=1363%2B1336_gateaway_transfection_on_9-14-2012" class="mw-redirect" title="1363+1336 gateaway transfection on 9-14-2012">1363+1336 gateaway transfection on 9-14-2012</a> to <a href="/index.php?title=363%2B1336_gateaway_transfection_on_9-14-2012" title="363+1336 gateaway transfection on 9-14-2012">363+1336 gateaway transfection on 9-14-2012</a> over redirect</p> <p><b>New page</b></p><div>[[Image:IMG 4237.JPG]]&lt;br&gt; <br /> <br /> {| width=&quot;532&quot; cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot;<br /> |-<br /> | No.&lt;br&gt; <br /> | &lt;br&gt; <br /> | ID&lt;br&gt; <br /> | Plasmid Name&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; &lt;br&gt; <br /> | miniprep <br /> | Lipofectanine 2000&lt;br&gt;<br /> |-<br /> | 1&lt;br&gt; <br /> | &lt;br&gt; <br /> | 393+1336-1&lt;br&gt; <br /> | pDonr221_EGFP+pCMV5-Fc(3C)-Dest-WPRE&lt;br&gt; <br /> | 15ul <br /> | weak green&lt;br&gt;<br /> |-<br /> | 2&lt;br&gt; <br /> | &lt;br&gt; <br /> | 393+1336-2&lt;br&gt; <br /> | pDonr221_EGFP+pCMV5-Fc(3C)-Dest-WPRE&lt;br&gt; <br /> | 15ul <br /> | weak green&lt;br&gt;<br /> |-<br /> | 3&lt;br&gt; <br /> | &lt;br&gt; <br /> | 393+1336-3&lt;br&gt; <br /> | pDonr221_EGFP+pCMV5-Fc(3C)-Dest-WPRE&lt;br&gt; <br /> | 15ul <br /> | weak green&lt;br&gt;<br /> |-<br /> | 5&lt;br&gt; <br /> | &lt;br&gt; <br /> | 393+1336-5 <br /> | pDonr221_EGFP+pCMV5-Fc(3C)-Dest-WPRE&lt;br&gt; <br /> | 10ul&lt;br&gt; <br /> | weak green <br /> | &lt;br&gt;<br /> |-<br /> | 6 <br /> | &lt;br&gt; <br /> | 393+1336-7 <br /> | pDonr221_EGFP+pCMV5-Fc(3C)-Dest-WPRE&lt;br&gt; <br /> | 10ul&lt;br&gt; <br /> | weak green&lt;br&gt; <br /> | &lt;br&gt;<br /> |-<br /> | 7 <br /> | &lt;br&gt; <br /> | 393+1336-8 <br /> | pDonr221_EGFP+pCMV5-Fc(3C)-Dest-WPRE&lt;br&gt; <br /> | 10ul&lt;br&gt; <br /> | weak green&lt;br&gt; <br /> | &lt;br&gt;<br /> |-<br /> | 8 <br /> | &lt;br&gt; <br /> | 529 <br /> | pLVX-mIL10-IRES-EGFP&lt;br&gt; <br /> | 0.5ul&lt;br&gt; <br /> | strong green&lt;br&gt; <br /> | &lt;br&gt;<br /> |}<br /> <br /> 12 well plate transfection (293T cells): <br /> <br /> &lt;br&gt; Purpose: want to check the gateaway plasmid expression in the 393 plus 1336 plasmid using 293T cells.&lt;br&gt; <br /> <br /> 1ug DNA plasmid (miniprep) and 4 ul lipofectamine.&amp;nbsp; Observed weak GFP at very late time 3rd day . For clone 529, transfection efficiency as expected <br /> <br /> [[Image:Gataaway-1 transfection.jpg]]&lt;br&gt; <br /> <br /> [[Image:Gateaway-2 transfection.jpg]]&lt;br&gt; <br /> <br /> Conclutsion: weak GFP expression with Lipofectamine 2000. Can&#039;t tell which plasmid has better expression.&lt;br&gt; <br /> <br /> [[Image:IMG 4237.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> [[Category:Cloning]] [[Category:QC]]</div> Paul.xu