https://eln.excellgen.com/index.php?action=history&feed=atom&title=1593_pT7A3-MBP-%283c%29-E_coli_Endo_IV 1593 pT7A3-MBP-(3c)-E coli Endo IV - Revision history 2026-10-18T11:28:28Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1593_pT7A3-MBP-(3c)-E_coli_Endo_IV&diff=8771&oldid=prev Paul.xu at 18:34, 1 April 2013 2013-04-01T18:34:18Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of ‎1593 pT7A3-MBP-(3c)-E coli Endo IV&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1593 pT7A3-MBP-(3c)-E coli Endo IV <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2013-2-7--- <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:1593&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039; Shanyi <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039;&lt;br&gt; ==<br /> <br /> === PCR ===<br /> <br /> PCR from genomic DNA isolated from Escherichia coli str. K-12 substr. MG1655 &lt;br&gt; <br /> <br /> [[Image:1572 1588 1593pcr.JPG]]&lt;br&gt; <br /> <br /> Cut out the band, digest PCR products and&amp;nbsp;1514 pT7A3-MBP-(3C) with Nde1 + Xho1&lt;br&gt; <br /> <br /> [[Image:DIGEST FOR 1593.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot; style=&quot;width: 281px; height: 66px;&quot;<br /> |-<br /> | &lt;br&gt; <br /> | band 1&lt;br&gt; <br /> | band 2&lt;br&gt;<br /> |-<br /> | &lt;br&gt; <br /> | pcr&lt;br&gt; <br /> | 1514&lt;br&gt;<br /> |-<br /> | enzymes&lt;br&gt; <br /> | colspan=&quot;2&quot; | &amp;nbsp;&amp;nbsp;&amp;nbsp; Nde1 + Xho1&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- <br /> <br /> Make miniprep and digest withNde1 + Xho1 <br /> <br /> [[Image:1593digest.JPG‎ ]]<br /> <br /> &lt;br&gt; <br /> <br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> == Protein Expression ==<br /> <br /> 2013/3/29&lt;br&gt; SHANYI <br /> <br /> [[Image:1097 1138 1458 1494 1505 1513 1558 1562 1573 1593 1654NEW.jpg]]&lt;br&gt; <br /> <br /> {| width=&quot;832&quot; cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot;<br /> |-<br /> | Lane&lt;br&gt; <br /> | Name&lt;br&gt; <br /> | Protein Name&lt;br&gt; <br /> | Clone ID&lt;br&gt; <br /> | Host&lt;br&gt; <br /> | Induction&lt;br&gt; <br /> | Purification&lt;br&gt; <br /> | pI/Mw&lt;br&gt;<br /> |-<br /> | M&lt;br&gt; <br /> | PageRuler Plus Prestained protein ladder&lt;br&gt; <br /> | &amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; &lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt;<br /> |-<br /> | 1&lt;br&gt; <br /> | pT7A3-T4-G32&lt;br&gt; <br /> | T4-G32&lt;br&gt; <br /> | 1097&lt;br&gt; <br /> | <br /> 5 ml,&lt;br&gt; <br /> <br /> Nico21(DE3)plysN <br /> <br /> | 0.2 mM IPTG&amp;nbsp; RT o/n&lt;br&gt; <br /> | Ni beads&lt;br&gt; <br /> | <br /> 4.82 / 33506.43 <br /> <br /> |-<br /> | 2&lt;br&gt; <br /> | pT7A3-MBP-(3c)-MutM&lt;br&gt; <br /> | MBP-(3c)-MutM&lt;br&gt; <br /> | 1138&lt;br&gt; <br /> | <br /> &amp;nbsp;5 ml,&lt;br&gt; Nico21(DE3)plysN&lt;br&gt; <br /> <br /> | 0.2 mM IPTG RT o/n &lt;br&gt; <br /> | Ni beads&lt;br&gt; <br /> | <br /> 8.68 / 30290.07+43.6 &lt;br&gt; <br /> <br /> |-<br /> | 3&lt;br&gt; <br /> | pT7A3-KOD&lt;br&gt; <br /> | KOD&lt;br&gt; <br /> | 1458&lt;br&gt; <br /> | <br /> &amp;nbsp; 5 ml,&lt;br&gt; Nico21(DE3)plysN&lt;br&gt; <br /> <br /> | 0.2 mM IPTG RT o/n &lt;br&gt; <br /> | Ni beads &lt;br&gt; <br /> | <br /> 7.53 / 90022.42 <br /> <br /> |-<br /> | 4&lt;br&gt; <br /> | pT7A3-UDG&lt;br&gt; <br /> | UDG&lt;br&gt; <br /> | 1494&lt;br&gt; <br /> | <br /> &amp;nbsp;5 ml,&lt;br&gt; Nico21(DE3)plysN&lt;br&gt; <br /> <br /> | 0.2 mM IPTG RT o/n &lt;br&gt; <br /> | Ni beads&lt;br&gt; <br /> | <br /> 5.45 / 27174.73 <br /> <br /> |-<br /> | 5&lt;br&gt; <br /> | pT7A3-T7-Endo1&lt;br&gt; <br /> | T7-Endo1&lt;br&gt; <br /> | 1505&lt;br&gt; <br /> | <br /> &amp;nbsp;5 ml,&lt;br&gt; Nico21(DE3)plysN&lt;br&gt; <br /> <br /> | 0.2 mM IPTG RT o/n &lt;br&gt; <br /> | Ni beads&lt;br&gt; <br /> | <br /> 9.54 / 17171.84 &amp;nbsp;&lt;br&gt; <br /> <br /> |-<br /> | 6&lt;br&gt; <br /> | pT7A3-polyAPol&lt;br&gt; <br /> | polyAPol&lt;br&gt; <br /> | 1513&lt;br&gt; <br /> | &amp;nbsp; 5 ml, <br /> Nico21(DE3)plysN&lt;br&gt; <br /> <br /> | 0.2 mM IPTG RT o/n &lt;br&gt; <br /> | Ni beads&lt;br&gt; <br /> | <br /> 9.67 / 53870.91&lt;br&gt; <br /> <br /> |-<br /> | 7&lt;br&gt; <br /> | pT7A3-T7-Exo&lt;br&gt; <br /> | T7-Exo&lt;br&gt; <br /> | 1558&lt;br&gt; <br /> | &amp;nbsp; 5 ml, <br /> Nico21(DE3)plysN&lt;br&gt; <br /> <br /> | 0.2 mM IPTG RT o/n &lt;br&gt; <br /> | Ni beads&lt;br&gt; <br /> | <br /> 4.93 / 34502.20<br /> <br /> |-<br /> | 8&lt;br&gt; <br /> | pT7A3-MBP-(3C)-Exo-1&lt;br&gt; <br /> | MBP-(3C)-Exo-1&lt;br&gt; <br /> | 1562&lt;br&gt; <br /> | &amp;nbsp; 5 ml, <br /> Nico21(DE3)plysN&lt;br&gt; <br /> <br /> | 0.2 mM IPTG RT o/n &lt;br&gt; <br /> | Ni beads&lt;br&gt; <br /> | <br /> 8.80 / 18143.72&lt;br&gt; <br /> <br /> |-<br /> | 9&lt;br&gt; <br /> | pT7A3-MBP-(3C)-RnaseH&lt;br&gt; <br /> | MBP-(3C)-RnaseH&lt;br&gt; <br /> | 1573&lt;br&gt; <br /> | &amp;nbsp; 5 ml, <br /> Nico21(DE3)plysN&lt;br&gt; <br /> <br /> | 0.2 mM IPTG RT o/n &lt;br&gt; <br /> | Ni beads&lt;br&gt; <br /> | <br /> 8.43 / &lt;br&gt; <br /> <br /> 17596.99 +43.6 <br /> <br /> |-<br /> | 10&lt;br&gt; <br /> | pT7A3-MBP-(3c)-E_coli_Endo IV&lt;br&gt; <br /> | MBP-(3c)-E_coli_Endo IV&lt;br&gt; <br /> | 1593&lt;br&gt; <br /> | &amp;nbsp; 5 ml, <br /> Nico21(DE3)plysN&lt;br&gt; <br /> <br /> | 0.2mM IPTG RT o/n &lt;br&gt; <br /> | Ni beads&lt;br&gt; <br /> | <br /> 5.43 / &lt;br&gt; <br /> <br /> 31479.53 +43.6 <br /> <br /> |-<br /> | 11&lt;br&gt; <br /> | pT7A3-T3 DNA ligase&lt;br&gt; <br /> | T3 DNA ligase&lt;br&gt; <br /> | 1654&lt;br&gt; <br /> | 5 ml, <br /> Nico21(DE3)plysN&lt;br&gt; <br /> <br /> | 0.2 mM IPTG RT o/n&lt;br&gt; <br /> | Ni beads&lt;br&gt; <br /> | 5.14 / 39350.70 &lt;br&gt;<br /> |}<br /> <br /> &lt;br&gt; <br /> <br /> <br /> [[Category:Cloning]]</div> Paul.xu