https://eln.excellgen.com/index.php?action=history&feed=atom&title=PAAV-SMN1_302 PAAV-SMN1 302 - Revision history 2026-10-18T08:46:31Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=PAAV-SMN1_302&diff=1309&oldid=prev Paul.xu at 17:31, 17 July 2011 2011-07-17T17:31:05Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of SMN1 to dsAAV-CB-MCS&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Gene synthesis of SMN1 and clone to dsAAV-CB-MCS . <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2011-7-12 <br /> <br /> &#039;&#039;&#039;Notebook: Haiyan-1&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:002995&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID:302&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039; Haiyan <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> [[Image:Smn1.JPG]] <br /> <br /> PCR amplify: primer SMN1-12; Pfu enzyme, 25 cycles; &lt;br&gt; <br /> <br /> Lane 1: Marker &lt;br&gt; <br /> <br /> Lane 2: 10 ul SMN1 pcr amplified product&lt;br&gt; <br /> <br /> Lane 3: dsAAV-CB-MCS, BglII+PstI&lt;br&gt; <br /> <br /> Lane 4: SMN1 PCR, BglII+PstI&lt;br&gt; <br /> <br /> Lane 5: SMN1 PCR, BglII+PstI&lt;br&gt; <br /> <br /> Ligate #3 with #4, #5 <br /> <br /> --[[User:Hylei|Hylei]] 17:29, 13 July 2011 (EST) <br /> <br /> == &#039;&#039;&#039;Verification&lt;br&gt;&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Restriction Digestion&#039;&#039;&#039;&lt;br&gt;<br /> <br /> &lt;br&gt;<br /> <br /> [[Image:PAAV_Cre_7_17_2011.JPG]]&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 &lt;br&gt;<br /> <br /> --[[User:Paul.xu|Paul.xu]] 12:31, 17 July 2011 (EST)<br /> <br /> &#039;&#039;&#039;Sequencing&#039;&#039;&#039;:&lt;br&gt;<br /> <br /> &lt;br&gt;<br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> *&amp;nbsp; pDs-AAV-SMN1 #1 and #2 may be good&lt;br&gt;<br /> *&amp;nbsp; <br /> *&amp;nbsp; <br /> *&amp;nbsp;&lt;br&gt;<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> &lt;br&gt; <br /> <br /> [[Category:Cloning]]</div> Paul.xu