https://eln.excellgen.com/index.php?action=history&feed=atom&title=PAAV-cre_256 PAAV-cre 256 - Revision history 2026-10-18T08:48:48Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=PAAV-cre_256&diff=1310&oldid=prev Paul.xu at 19:15, 17 July 2011 2011-07-17T19:15:34Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;SubCloning of Cre&amp;nbsp; to pAAV-MCS&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Subcloning of Cre to pAAV-MCS&lt;br&gt; <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2011-7-? to 2011-7-? <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/Paul <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === Subcloning ===<br /> <br /> Date: 2011-6-8 <br /> <br /> Digest pLVGP-cre (clone #236) with Ecori + XhoI&lt;br&gt; <br /> <br /> Digest pAAV-MCS (clone #241) with Ecori + XhoI&lt;br&gt; <br /> <br /> cut out insert and vector, ligate and transform STBL3 <br /> <br /> [[Image:AAV.JPG]] <br /> <br /> Lane 1: Marker &lt;br&gt; <br /> <br /> Lane 2: pLVGP-cre , EcoR1 + XhoI&lt;br&gt; <br /> <br /> Lane 3: pAAV-MCS, EcoRI + XhoI&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Colony Screening&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Digestion&#039;&#039;&#039; <br /> <br /> [[Image:PAAV Cre 7 17 2011.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> Lane 1: pAAV-Cre #1, digest with EcoR1 + Xho1 <br /> <br /> Lane 2: pAAV-Cre #2, digest with EcoR1 + Xho1 <br /> <br /> Lane 3: pAAV-Cre #3, digest with EcoR1 + Xho1 <br /> <br /> Lane 4: pDs-AAV-SMN1 #1, digest with BglII + Pst1 <br /> <br /> Lane 5: pDs-AAV-SMN1 #2, digest with BglII + Pst1 <br /> <br /> Lane 6: pDs-AAV-SMN1 #3, digest with BglII + Pst1 <br /> <br /> Lane 7: pAAV-Cre #1, digest with Nco1 + Xho1 <br /> <br /> Lane 8: pAAV-Cre #2, digest with Nco1 + Xho1 <br /> <br /> Lane 9: pAAV-Cre #3, digest with Nco1 + Xho, well leaks <br /> <br /> Lane 10: pET30b , digest with Nde1 + Nco1 <br /> <br /> --[[User:Paul.xu|Paul.xu]] 12:15, 17 July 2011 (EST) <br /> <br /> &#039;&#039;&#039;Sequencing&#039;&#039;&#039;&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> == Result intepretation/Summary ==<br /> <br /> Looks like pAAV-Cre #2 &amp;amp; #3 are good. There is one Nco1 site in the CMV promoter&lt;br&gt; <br /> <br /> == Conclusion ==<br /> <br /> &lt;br&gt; <br /> <br /> == Comments ==<br /> <br /> ----<br /> <br /> [[Category:Cloning]]</div> Paul.xu