https://eln.excellgen.com/index.php?action=history&feed=atom&title=1290_pET28b-PTD-DRBD 1290 pET28b-PTD-DRBD - Revision history 2026-10-18T02:20:04Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1290_pET28b-PTD-DRBD&diff=7021&oldid=prev Paul.xu at 18:51, 14 November 2012 2012-11-14T18:51:01Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1290 pET28b-PTD-DRBD&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1290 pET28b-PTD-DRBD <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;August 1, 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;1290 <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 1, 2012&lt;br&gt; <br /> <br /> [[Image:1290 Amp.JPG]]&amp;lt;br&amp;gt; [[Image&amp;lt;br&amp;gt;&amp;nbsp;:1292 1307DIGEST10 31 2012.JPG]]&amp;lt;br&amp;gt; &lt;br&gt; <br /> <br /> Synthesized template via gene synthesis&lt;br&gt; <br /> <br /> Expected size ~400 bp (see lane 1)&lt;br&gt; <br /> <br /> Cut out band and purified&lt;br&gt; <br /> <br /> [[Image:1290 Digestion.JPG]]&lt;br&gt; <br /> <br /> Digested the insert (lane 1)&amp;nbsp;and the vector (V12, pET28b) (lane 2)&amp;nbsp;with RE N&amp;lt;br&amp;gt; de1 + Xho1&lt;br&gt; <br /> <br /> Cut out bands, pu&lt;br&gt; rify and ligate together overnight <br /> <br /> Date: August 2, 2012&lt;br&gt; <br /> <br /> Ligation went wrong yesterday, start all over&lt;br&gt; <br /> <br /> [[Image:1290 0802 Amp.JPG]]&lt;br&gt; <br /> <br /> Synthesized template via gene synthesis <br /> <br /> Expected size ~400 bp (see lane 1) <br /> <br /> Cut out band and purified&lt;br&gt; <br /> <br /> [[Image:1290 0802 Digestion.JPG]]&amp;lt;br&amp;gt; &lt;br&gt; <br /> <br /> Digested the insert (lane 1) and the vector (V12, pET28b) (lane 2) with RE Nde1 + Xho1 <br /> <br /> Cut out bands, purify and ligate together overnight &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> Date:&amp;nbsp;August 13, 2012&lt;br&gt; <br /> <br /> Sequencing came back with a lot of mutations&lt;br&gt; <br /> <br /> [[Image:1290 0813 Amp.JPG]] <br /> <br /> Synthesized template via gene synthesis&lt;br&gt; <br /> <br /> Expected size ~400 bp (see lane 1) <br /> <br /> Cut out band and purified&lt;br&gt; <br /> <br /> [[Image:1290 0813 Digestion.JPG]]&amp;lt;br&amp;gt; &lt;br&gt; <br /> <br /> Digested the insert (lane 1) and the vector (V12, pET28b) (lane 2) with RE Nde1 + Xho1 <br /> <br /> Cut out bands, purify and ligate together overnight &lt;br&gt; <br /> <br /> === &lt;u&gt;&lt;/u&gt;Colony Screening&lt;br&gt; ===<br /> <br /> Date:&amp;nbsp;August 7, 2012&lt;br&gt; <br /> <br /> Digested MiniPrep with RE&amp;nbsp;Nde1 and Xho1&lt;br&gt; <br /> <br /> Expected size ~400 bp&lt;br&gt; <br /> <br /> [[Image:1290 0807 Screening.JPG]] <br /> <br /> Samples 1, 2, 3, 5 and 6 may be correct <br /> <br /> Sent sample 5 to sequencing <br /> <br /> &lt;br&gt; <br /> <br /> Date: August 16, 2012 <br /> <br /> Digested MiniPrep with RE Nde1 and Xho1 <br /> <br /> Expected size ~400 bp&lt;br&gt; <br /> <br /> [[Image:1290 0816 Screening.JPG]] <br /> <br /> Samples 1, 2 and 6 may be correct. Send sample 6 to sequencing <br /> <br /> &lt;br&gt; &lt;br&gt; Date:&amp;nbsp;OCO 26, <br /> <br /> 2012 Digested insert&amp;nbsp; and&amp;nbsp; vector&amp;nbsp; with&amp;nbsp; Nde1 + Xho1&lt;br&gt; <br /> <br /> [[Image:1290V15 1292368 1293V15 1352 13041280DIGEST 10 26 2012.JPG]]&lt;br&gt; <br /> <br /> Ligate , transform&amp;nbsp; <br /> <br /> === &lt;u&gt;&lt;/u&gt;Colony Screening&lt;br&gt; ===<br /> <br /> [[Image:1304 1290 DIGEST 10 31 2012.JPG]] <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> Paul.xu