https://eln.excellgen.com/index.php?action=history&feed=atom&title=1800-pCold-6His-TEV-MBP 1800-pCold-6His-TEV-MBP - Revision history 2026-10-18T14:13:10Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1800-pCold-6His-TEV-MBP&diff=9861&oldid=prev Paul.xu at 20:19, 26 July 2013 2013-07-26T20:19:35Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1800-pCold-6His-TEV-MBP&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1800-pCold-6His-TEV-MBP <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2013-7-22---2013-7-24 <br /> <br /> &#039;&#039;&#039;Notebook:0&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID:1800&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039; yingying <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === cloning ===<br /> <br /> [[Image:1797p2+1780pcr 7-22-2013.JPG]]&lt;br&gt; <br /> <br /> 1. PCR TEV from 432 pMBP-7xHis-TEV <br /> <br /> TEV-Kpn1-F &lt;br&gt;TEV-MBP-R&lt;br&gt; <br /> <br /> 2,PCR MBP from 1281 pMAL-p5E as template <br /> <br /> TEV-MBP-F <br /> <br /> MBP-Sal1-stop-R2 &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> [[Image:1797pcr2+1798PCR+1800PCR 7-23-2013.JPG]]&lt;br&gt; <br /> <br /> 3. Join two fragments using &lt;br&gt;TEV-Kpn1-F &lt;br&gt;MBP-Sal1-stop-R2 &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> [[Image:1798+1800clone 7-23-2013.JPG]]&lt;br&gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot; width=&quot;326&quot;<br /> |-<br /> | lane&lt;br&gt; <br /> | 3&lt;br&gt; <br /> | 4&lt;br&gt;<br /> |-<br /> | &lt;br&gt; <br /> | PCR product&lt;br&gt; <br /> | v19 pET28-cold&lt;br&gt;<br /> |-<br /> | enzymes&lt;br&gt; <br /> | Kpn1 + Sal1&lt;br&gt; <br /> | Kpn1 + Xho1&lt;br&gt;<br /> |}<br /> <br /> &lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2013-7-24 <br /> <br /> Make miniprep and digest&amp;nbsp; Kpn1+Bgl11&lt;br&gt; <br /> <br /> [[Image:1800digest 7-24-2013.JPG]] <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Cloning of 1800-pCold-6His-TEV-MBP&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1800-pCold-6His-TEV-MBP <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2013-7-24--2013-7-26<br /> <br /> &#039;&#039;&#039;Notebook:0&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID:1800&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039; yingying <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === cloning ===<br /> <br /> [[Image:1797p2+1780pcr 7-22-2013.JPG]]&lt;br&gt; <br /> <br /> 1. PCR TEV from 432 pMBP-7xHis-TEV <br /> <br /> TEV-Kpn1-F &lt;br&gt;TEV-MBP-R&lt;br&gt; <br /> <br /> 2,PCR MBP from 1281 pMAL-p5E as template <br /> <br /> TEV-MBP-F <br /> <br /> MBP-Sal1-stop-R2 &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> [[Image:1800PCR 7-25-2013.JPG]] <br /> <br /> &lt;br&gt; <br /> <br /> 3. Join two fragments using &lt;br&gt;TEV-Kpn1-F &lt;br&gt;MBP-Sal1-stop-R2 &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> [[Image:1800clone 7-25-2013.JPG]]&lt;br&gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot; width=&quot;326&quot;<br /> |-<br /> | lane&lt;br&gt; <br /> | 3&lt;br&gt; <br /> | 4&lt;br&gt;<br /> |-<br /> | &lt;br&gt; <br /> | PCR product&lt;br&gt; <br /> | v19 pET28-cold&lt;br&gt;<br /> |-<br /> | enzymes&lt;br&gt; <br /> | Kpn1 + Sal1&lt;br&gt; <br /> | Kpn1 + Xho1&lt;br&gt;<br /> |}<br /> <br /> &lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2013-7-26<br /> <br /> Make miniprep and digest Kpn1 + Bgl11&lt;br&gt; <br /> <br /> [[Image:1800digest_7-26-2013.JPG]]<br /> <br /> clone1,3,4 may be right ,need to sequencing<br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> &lt;br&gt; <br /> <br /> [[Category:Cloning]]</div> Paul.xu