https://eln.excellgen.com/index.php?action=history&feed=atom&title=1193_pCDNA3.2-Fc 1193 pCDNA3.2-Fc - Revision history 2026-10-18T10:26:09Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1193_pCDNA3.2-Fc&diff=4328&oldid=prev Paul.xu at 17:09, 29 May 2012 2012-05-29T17:09:55Z <p></p> <p><b>New page</b></p><div>= &#039;&#039;&#039;pCDNA3.2-Fc (1193)&#039;&#039;&#039;&lt;br&gt; =<br /> <br /> &#039;&#039;&#039;Date: 2012-05-26&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Notebook:&#039;&#039;&#039;&amp;nbsp; 4480&lt;br&gt; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039;&amp;nbsp; 4481&lt;br&gt; <br /> <br /> &#039;&#039;&#039;Clone ID:&#039;&#039;&#039; 1193 <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039; &lt;span style=&quot;font-weight: bold;&quot;&gt;Paul&lt;/span&gt;&lt;br&gt; <br /> <br /> == Cloning ==<br /> <br /> PCR Fc domain from pFUSE-hIgG1-Fc2-IAB 1101 using these oligos: &lt;br&gt;ssFc-BamH1-F1 + 10% ss-Fc-F5 + Fc-Kpn1-R4, template is pFUSE-hIgG1-Fc2-IAB, 1101&lt;br&gt;ssFc-BamH1-F1&lt;br&gt;accgtcGGATCCGCCACCATGGGAGTCAAAGTTCTGTTTGCCCTGATCTGCATCGCTGTG&lt;br&gt; GGATCCGCCACCATGGGAGTCAAAGTTCTGTTTGCCCTGATCTGCATCGCTGTG GCC GAG GCC AAG CCC ACC&lt;br&gt;ss-Fc-F5 &lt;br&gt; CCCTGATCTGCATCGCTGTGGCCGAGGCCAAGCCCACCgacaaaactcacacatgcccac&lt;br&gt;Fc-Kpn1-R4&lt;br&gt;(ctctccctgtctccgggtaaaGGCAGCGGCGAGAACCTGTACTTCCAGGGCggtaccATCGAC)&lt;br&gt;tactagggtaccGCCCTGGAAGTACAGGTTCTCGCCGCTGCCtttacccggagacagggagag <br /> <br /> Digest with BamH1 + Kpn1, clone into pCDNA3.2, 1192&lt;br&gt; <br /> <br /> [[Image:1193 1198 1199 PCR 5 26 2012.jpg]]&lt;br&gt; <br /> <br /> 5/27/2012&lt;br&gt; <br /> <br /> digest PCR product and vector with BamH1 + Kpn1&lt;br&gt; <br /> <br /> [[Image:1193 1198 1199 cloning PCR frag 5 27 2012.jpg]]&lt;br&gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot; style=&quot;width: 298px; height: 106px;&quot;<br /> |-<br /> | 1&lt;br&gt; <br /> | 2&lt;br&gt; <br /> | 3&lt;br&gt; <br /> | 4&lt;br&gt;<br /> |-<br /> | 1192 pCDNA3.2&lt;br&gt; <br /> | <br /> 1198 <br /> <br /> ssRFP-Fc-(3c)&lt;br&gt; <br /> <br /> | <br /> 1193 <br /> <br /> Fc&lt;br&gt; <br /> <br /> | <br /> 1199 <br /> <br /> ssRFP-Fc-(tev)&lt;br&gt; <br /> <br /> |}<br /> <br /> gel purified, and ligate, transform DH5alpha.&lt;br&gt; <br /> <br /> == Colony Screening by Restriction Digestion&lt;br&gt; ==<br /> <br /> 5-29-2012<br /> <br /> [[Image:1193_digest_detect_5-29-2012.JPG]]<br /> <br /> clone 2,4,6 may be right, neeed to sequence<br /> <br /> &lt;br&gt; <br /> <br /> [[Category:Cloning]]</div> Paul.xu