https://eln.excellgen.com/index.php?action=history&feed=atom&title=1353-1305-1369_transfection_on_0914-2012 1353-1305-1369 transfection on 0914-2012 - Revision history 2026-10-18T05:56:15Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1353-1305-1369_transfection_on_0914-2012&diff=6193&oldid=prev Paul.xu at 20:48, 23 September 2012 2012-09-23T20:48:18Z <p></p> <p><b>New page</b></p><div>[[Image:IMG 4237.JPG]]&lt;br&gt; <br /> <br /> {| width=&quot;434&quot; cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot;<br /> |-<br /> | No.&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 /> | conc.(ug/ul) <br /> | Lipofectanine 2000&lt;br&gt;<br /> |-<br /> | 1&lt;br&gt; <br /> | 1353&lt;br&gt; <br /> | pCMV5-Fc(3C)-RFP-WPRE&lt;br&gt; <br /> | 2.0 <br /> | weak red at 50h&lt;br&gt;<br /> |-<br /> | 2&lt;br&gt; <br /> | 1305&lt;br&gt; <br /> | pCMV5-Fc(3C)-TagRFP&lt;br&gt; <br /> | 1.0 <br /> | weak red at 50h&lt;br&gt;<br /> |-<br /> | 3&lt;br&gt; <br /> | 1369&lt;br&gt; <br /> | pCMV5-Fc-(3c)-TagRFP-WPRE-oriP&lt;br&gt; <br /> | 2.8 <br /> | weak red at 50h&lt;br&gt;<br /> |-<br /> | 5&lt;br&gt; <br /> | 529&lt;br&gt; <br /> | pLVX-mIL10-IRES-EGFP&lt;br&gt; <br /> | 2.8 <br /> | strong&lt;br&gt;<br /> |}<br /> <br /> 12 well plate transfection (293T cells):&lt;br&gt; Purpose: want to check the stongest expression in the 1353, 1305, 1369 plasmid using 293T cells.&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> To avoid totoxity of lipofectamine,&amp;nbsp; use 0.5ug DNA plasmid (made 1:20 dilution) and 2 ul lipofectamine. Change medium after 5 hs transfection. Observed weak RFP at 50 hs on monday. For clone 529, transfection efficiency as expected and 0.5 ug plasmid is less efficient than 1 ug plasmid.&lt;br&gt; <br /> <br /> [[Image:Transfection 9 19 2012.jpg]] <br /> <br /> 9/21/2012&lt;br&gt; <br /> <br /> [[Image:1369 1353 1305 2012 9 21.jpg]]&lt;br&gt; <br /> <br /> 1369: pCMV5-Fc-(3c)-TagRFP-WPRE-oriP&lt;br&gt; <br /> <br /> 1353: pCMV5-Fc(3C)-RFP-WPRE &lt;br&gt; <br /> <br /> 1305 pCMV5-Fc(3C)-TagRFP&lt;br&gt; --[[User:Paul.xu|Paul.xu]] 17:25, 21 September 2012 (EST) &lt;br&gt; <br /> <br /> Photo taken on 9/23/2012 <br /> <br /> [[Image:1369_1353_1305_2012_9_23.jpg]]<br /> <br /> [[Image:9 23 2012 2.jpg]] <br /> <br /> [[Image:9 23 2012 1.jpg]] <br /> <br /> Conclutsion: weak RFP 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