https://eln.excellgen.com/index.php?action=history&feed=atom&title=1764-pT7A3-T4-Pnk 1764-pT7A3-T4-Pnk - Revision history 2026-10-18T05:02:26Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1764-pT7A3-T4-Pnk&diff=9706&oldid=prev Paul.xu at 14:54, 12 July 2013 2013-07-12T14:54:33Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1764-pT7A3-T4-Pnk&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1764-pT7A3-T4-Pnk <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2013-4-1----2013-4-5 <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:1764&#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:1704 PCR 4-1-2013.JPG]] <br /> <br /> [[Image:1704AB clone 4-2-2013.JPG]] <br /> <br /> A&amp;nbsp;: vector:PET28b <br /> <br /> B: vector 1454 pEC <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2013-4 <br /> <br /> Make miniprep and digest with Nde1+xho1&lt;br&gt; <br /> <br /> [[Image:1704 digest detect 4-5-2013.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 /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1764-pT7A3-T4-Pnk <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2013-6-3---2013-6-7 <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:1764&#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 /> &lt;br&gt; <br /> <br /> [[Image:1704PCR 5-31-2013.JPG]]&lt;br&gt; <br /> <br /> [[Image:1619+1704+1709clone 6-3-2013.JPG]] <br /> <br /> vector:1279-pT7A3 <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2013-6-7 <br /> <br /> Make miniprep and digest Nde1+Xho1&lt;br&gt; <br /> <br /> [[Image:556+1704 digest 6-7-2013.JPG]] <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 /> === &#039;&#039;&#039;protein screening&#039;&#039;&#039; ===<br /> <br /> [[Image:643+645+652+1721+1731+1736+1757+1764+1771protein_screening7-9-2013.jpg]]<br /> <br /> Ni beads <br /> <br /> {| width=&quot;200&quot; cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot;<br /> |-<br /> | lane&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 /> | colspan=&quot;2&quot; | MW&lt;br&gt; <br /> | Conclusion&lt;br&gt;<br /> |-<br /> | 1&lt;br&gt; <br /> | 643,pT7A3-Nhe1&lt;br&gt; <br /> | 4ml,NiCO21(DE3)pLysN&lt;br&gt; <br /> | 0.2 mM IPTG 25 oC o/n&lt;br&gt; <br /> | Ni beads&lt;br&gt; <br /> | colspan=&quot;2&quot; | 38KD&lt;br&gt; <br /> | yes&lt;br&gt;<br /> |-<br /> | 2&lt;br&gt; <br /> | 645,pT7A3-Pst1&lt;br&gt; <br /> | 4ml,NiCO21(DE3)pLysN&lt;br&gt; <br /> | 0.2 mM IPTG 25 oC o/n&lt;br&gt; <br /> | Ni beads&lt;br&gt; <br /> | colspan=&quot;2&quot; | 37KD&lt;br&gt; <br /> | weak&lt;br&gt;<br /> |-<br /> | 3&lt;br&gt; <br /> | 652, pT7A30-hIFNA2&lt;br&gt; <br /> | 4 ml, <br /> BL21(DE3)Star, pRare2-pLysS &lt;br&gt; <br /> <br /> | 0.2 mM IPTG 25 oC o/n&lt;br&gt; <br /> | Ni beads&lt;br&gt; <br /> | 20KD weak&lt;br&gt; <br /> | <br /> |-<br /> | 4&lt;br&gt; <br /> | 1721, pT7A3-MBP-(E3C)-SOD&lt;br&gt; <br /> | 4ml,NiCO21(DE3)pLysN&lt;br&gt; <br /> | 0.2 mM IPTG 25 oC o/n&lt;br&gt; <br /> | Ni beads&lt;br&gt; <br /> | colspan=&quot;2&quot; | 60KD&lt;br&gt; <br /> | only MBP&lt;br&gt;<br /> |-<br /> | 5&lt;br&gt; <br /> | 1731,pT7A3-TurboEnterokinase&lt;br&gt; <br /> | 4 ml, <br /> BL21(DE3)Star, pRare2-pLysS &lt;br&gt; <br /> <br /> | 0.2 mM IPTG 25 oC o/n&lt;br&gt; <br /> | Ni beads&lt;br&gt; <br /> | colspan=&quot;2&quot; | 27KD&lt;br&gt; <br /> | &amp;nbsp;?&lt;br&gt;<br /> |-<br /> | 6&lt;br&gt; <br /> | 1736,pT7A3-His-NusA&lt;br&gt; <br /> | 4 ml, <br /> BL21(DE3)Star, pRare2-pLysS &lt;br&gt; <br /> <br /> | 0.2 mM IPTG 25 oC o/n&lt;br&gt; <br /> | Ni beads&lt;br&gt; <br /> | colspan=&quot;2&quot; | 56KD&lt;br&gt; <br /> | yes&lt;br&gt;<br /> |-<br /> | 7&lt;br&gt; <br /> | 1757,pT7A3-MBP-(3C)-FN3K&lt;br&gt; <br /> | 4 ml, <br /> BL21(DE3)Star, pRare2-pLysS &lt;br&gt; <br /> <br /> | 0.2 mM IPTG 25 oC o/n&lt;br&gt; <br /> | Ni beads&lt;br&gt; <br /> | colspan=&quot;2&quot; | 81KD&lt;br&gt; <br /> | weak&lt;br&gt;<br /> |-<br /> | 8&lt;br&gt; <br /> | 1764,pT7A3-T4-Pnk&lt;br&gt; <br /> | 4ml,NiCO21(DE3)pLysN&lt;br&gt; <br /> | 0.2 mM IPTG 25 oC o/n&lt;br&gt; <br /> | Ni beads&lt;br&gt; <br /> | colspan=&quot;2&quot; | 37KD&lt;br&gt; <br /> | yes&lt;br&gt;<br /> |-<br /> | 9&lt;br&gt; <br /> | 1771,pT7A3-MBP-(3C)-PCK1&lt;br&gt; <br /> | 4 ml, <br /> BL21(DE3)Star, pRare2-pLysS &lt;br&gt; <br /> <br /> | 0.2 mM IPTG 25 oC o/n&lt;br&gt; <br /> | Ni beads&lt;br&gt; <br /> | colspan=&quot;2&quot; | 120KD&lt;br&gt; <br /> | yes&lt;br&gt;<br /> |}<br /> <br /> &lt;br&gt; <br /> <br /> <br /> [[Category:Cloning]]</div> Paul.xu