https://eln.excellgen.com/index.php?action=history&feed=atom&title=1251_pET28b-KOD-sso7d 1251 pET28b-KOD-sso7d - Revision history 2026-10-18T04:25:37Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1251_pET28b-KOD-sso7d&diff=5185&oldid=prev Xiaojing.Wang at 15:46, 19 July 2012 2012-07-19T15:46:37Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1251 pET28b-KOD-sso7d&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1251 pET28b-KOD-sso7d <br /> <br /> &#039;&#039;&#039;Date:&#039;&#039;&#039; July 12, 2012&#039;&#039;&#039;&lt;br&gt;&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Notebook:&amp;nbsp;&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID: &#039;&#039;&#039;1251 <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039; Jennifer Truong&lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === Cloning ===<br /> <br /> Date: July 12, 2012&lt;br&gt; <br /> <br /> [[Image:1251 digestion.JPG]]&lt;br&gt; <br /> <br /> Digested 1250 pET28b-KOD-2 with Nde1 + Kpn1 --&amp;gt; insert (lane 1, bottom band)&lt;br&gt;Digested 368 pET30b-sso7d-KX with Nde1 + Kpn1 --&amp;gt;vector (lane 2, top band)<br /> <br /> &lt;br&gt;Date:&amp;nbsp;July 17, 2012&lt;br&gt; <br /> <br /> [[Image:1251_0717_digestion2.JPG]]<br /> <br /> MiniPrep digestion was strange, digest insert and vector again. Only two bands this time, instead of three. Do not know why there were three bands before, maybe used the wrong enzymes. Ligate together <br /> <br /> Digested 1250 pET28b-KOD-2 with Nde1 + Kpn1 --&amp;gt; insert (lane 1, bottom band)&lt;br&gt;Digested 368 pET30b-sso7d-KX with Nde1 + Kpn1 --&amp;gt;vector (lane 2, top band)&lt;br&gt;<br /> <br /> Cut out band from gel, purify, ligated for 1 hour and transform with Top10 cells and plated on Kan plates to grow over night <br /> <br /> === Colony Screening ===<br /> <br /> Date:&amp;nbsp;July 16, 2012&lt;br&gt; <br /> <br /> [[Image:1251 0716 Screening.JPG]]&lt;br&gt; <br /> <br /> Digested MiniPrep with Nde1 + Kpn1 &lt;br&gt; <br /> <br /> July 17, 2012 <br /> <br /> [[Image:1251 0717 digestion.JPG]] <br /> <br /> Digested MiniPrep again with Nde1 +&amp;nbsp;Kpn1 a little bit longer. Maybe there was something wrong with the digested before, should ligate together and transform again <br /> <br /> Date:&amp;nbsp;July 19, 2012&lt;br&gt;<br /> <br /> [[Image:1151_0719_Screening.JPG]]&lt;br&gt;<br /> <br /> Digested MiniPrep again with Nde1 + Kpn1 for 1 hr 15 minutes&lt;br&gt;<br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039;&lt;br&gt; ==<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> &lt;br&gt; <br /> <br /> [[Category:Cloning]]</div> Xiaojing.Wang