https://eln.excellgen.com/index.php?action=history&feed=atom&title=1297_pET-6H-VPMLK-TAT-NLS 1297 pET-6H-VPMLK-TAT-NLS - Revision history 2026-10-18T02:19:40Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1297_pET-6H-VPMLK-TAT-NLS&diff=5701&oldid=prev Jennifer.truong at 13:42, 21 August 2012 2012-08-21T13:42:46Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1297 pET-6H-VPMLK-TAT-NLS&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1297 pET-6H-VPMLK-TAT-NLS <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;August 13, 2012&lt;br&gt; <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;1297 <br /> <br /> &#039;&#039;&#039;Experimenters: &#039;&#039;&#039;Jennifer Truong <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === Cloning&lt;br&gt; ===<br /> <br /> Date:&amp;nbsp;August 13, 2012\ <br /> <br /> Gene synthesis <br /> <br /> Purify PCR product, see lane 2, (113 bp, need to add 10% isopropanol becasue the fragment is very small). <br /> <br /> [[Image:1297 0813 Amp.JPG]] <br /> <br /> Digest PCR product with RE Nco1 + Nde1.&lt;br&gt;Digest vector 1128 pDB.His.PfuMBP-(tev) with Nco1 + Nde1.&lt;br&gt;Ligate together overnight and transform&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> Date:&amp;nbsp;August 14, 2012 <br /> <br /> Insert disappeared after digestion, need to reamplify, see lane 1 <br /> <br /> [[Image:0814 Amp.JPG]] <br /> <br /> Date:&amp;nbsp;August 15, 2012&lt;br&gt; <br /> <br /> [[Image:1297 0815 Digestion.JPG]]&lt;br&gt; <br /> <br /> Digest PCR product with RE Nco1 + Nde1 (lane 1)&lt;br&gt;Digest vector pDB.His.MBP, V14 with Nco1 + Nde1 (lane 2)&lt;br&gt;Ligate together and transform&lt;br&gt; <br /> <br /> === &lt;u&gt;&lt;/u&gt;Colony Screening&lt;br&gt; ===<br /> <br /> Date:&amp;nbsp;August 20, 2012&lt;br&gt; <br /> <br /> [[Image:1297 Screening.JPG]] <br /> <br /> Digested MiniPrep with RE&amp;nbsp;Nco1 and Nde1<br /> <br /> Faint band at expected size (~100 bp)&amp;nbsp;maybe send to sequencing <br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> &lt;br&gt; <br /> <br /> [[Category:Cloning]]</div> Jennifer.truong