https://eln.excellgen.com/index.php?action=history&feed=atom&title=1035-pET28b-Tth-Ligase-K294R 1035-pET28b-Tth-Ligase-K294R - Revision history 2026-10-18T05:51:47Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1035-pET28b-Tth-Ligase-K294R&diff=9804&oldid=prev Paul.xu at 19:10, 23 July 2013 2013-07-23T19:10:51Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1035-pET28b-Tth-Ligase-K294R&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1035-pET28b-Tth-Ligase-K294R <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2012-5-10----2012-5-17 <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:1035&#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:1035 PCR1+2 5-14-2012.JPG]][[Image:1035 PCR 5-14-2012.JPG]]&lt;br&gt; <br /> <br /> PCR using &lt;br&gt; <br /> <br /> 1stPCR:pET28b-Tth-Ligase 1022 as template, Tth-Lig-Nde1-F + TagLiase-K294R-R <br /> <br /> 2ndPCR: pET28b-Tth-Ligase 1022 as template, TagLiase-K294R-F + TagLiase-BamH1-R&lt;br&gt; <br /> <br /> 3rdPCR 1stPCR+2ndPCR as template, Tth-Lig-Nde1-F +TagLiase-BamH1-R&lt;br&gt; <br /> <br /> [[Image:537+1035+1062+1063clone 5-15-2012.JPG]]&lt;br&gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot; style=&quot;width: 446px; height: 84px;&quot;<br /> |-<br /> | &lt;br&gt; <br /> | band 2&lt;br&gt; <br /> | band 3&lt;br&gt;<br /> |-<br /> | &lt;br&gt; <br /> | PCR product&lt;br&gt; <br /> | pET28b-Tth-Ligase 1022&lt;br&gt;<br /> |-<br /> | enzymes&lt;br&gt; <br /> | colspan=&quot;2&quot; | &amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; Nde1 + BamH1 &lt;br&gt;<br /> |}<br /> <br /> &lt;br&gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot;<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> === Colony Screening ===<br /> <br /> Date: 2012-5-17 <br /> <br /> Make miniprep and digest with Nde1 + BamH1 <br /> <br /> [[Image:1035 digest detect 5-17-2012.JPG]] <br /> <br /> clone 8 may be right, need to sequence <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> *&amp;amp;lt;br&amp;amp;gt;<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> &amp;amp;lt;br&amp;amp;gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot;<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> &amp;amp;lt;br&amp;amp;gt; <br /> <br /> {| cellspacing=&quot;1&quot;<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> &amp;lt;br&amp;gt; <br /> <br /> {| cellspacing=&quot;1&quot;<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Cloning of 1035-pET28b-Tth-Ligase-K294R&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1035-pET28b-Tth-Ligase-K294R <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2013-7-9--- <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:1035&#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:1035pcr1+2_7-8-2013.JPG]]&lt;br&gt; <br /> <br /> PCR using &lt;br&gt; <br /> <br /> 1stPCR:pET28b-Tth-Ligase 1022 as template, Tth-Lig-Nde1-F + TagLiase-K294R-R <br /> <br /> 2ndPCR: pET28b-Tth-Ligase 1022 as template, TagLiase-K294R-F + TagLiase-BamH1-R&lt;br&gt; <br /> <br /> <br /> <br /> [[Image:1035PCR+1780PCR2_7-17-2013.jpg]]<br /> <br /> 3rdPCR 1stPCR+2ndPCR as template, Tth-Lig-Nde1-F +TagLiase-BamH1-R&lt;br&gt; <br /> <br /> <br /> <br /> [[Image:1035_clone_7-18-2013.JPG]]<br /> <br /> &lt;br&gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot; style=&quot;width: 446px; height: 84px;&quot;<br /> |-<br /> | &lt;br&gt; <br /> | band 2&lt;br&gt; <br /> | band 3&lt;br&gt;<br /> |-<br /> | &lt;br&gt; <br /> | PCR product&lt;br&gt; <br /> | pET28b-Tth-Ligase 1022&lt;br&gt;<br /> |-<br /> | enzymes&lt;br&gt; <br /> | colspan=&quot;2&quot; | &amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; Nde1 + BamH1 &lt;br&gt;<br /> |}<br /> <br /> &lt;br&gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot;<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> === Colony Screening ===<br /> <br /> Date: 2013-7-23 <br /> <br /> Make miniprep and digest with Nde1 + BamH1 <br /> <br /> [[Image:1035+1780digest_7-23-2013.JPG]]&lt;br&gt; <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 /> <br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> <br /> <br /> [[Category:Cloning]]</div> Paul.xu