https://eln.excellgen.com/index.php?action=history&feed=atom&title=1217_pDB.His.PfuMBP-%283C%29 2026-10-18T02:19:36Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1217_pDB.His.PfuMBP-(3C)&diff=5700&oldid=prev 2012-08-21T13:15:21Z

<p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1217 pDB.His.PfuMBP-(3C)&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1217 pDB.His.PfuMBP-(3C) <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;June 26, 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: 1217&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters: &#039;&#039;&#039;Jennifer Truong <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === Cloning ===<br /> <br /> [[Image:0626 Amp.JPG]]&lt;br&gt; <br /> <br /> Digested pDB.His.PfuMBP 1129 with Nde1 + BamH1 (see lane 1), looks like it is not digested. Ran out of 1129 vector, growing new from gycerol stock. Will redo tomorrow. Stored annealed oligos in box.&lt;br&gt; <br /> <br /> Date:&amp;nbsp;June 27, 2012&lt;br&gt; <br /> <br /> Digested pDB.His.PfuMBP 1129 with Nde1 + BamH1 (see lane 1) again, same 2 bands happened. &lt;br&gt; <br /> <br /> [[Image:0627 Digestion.JPG]] <br /> <br /> Digested vector separately with BamHI and Nde1, digest with NEB enzymes&lt;br&gt; <br /> <br /> [[Image:1192 digestion.JPG]] <br /> <br /> Digested with NEB BamHI and Nde1 <br /> <br /> [[Image:1217 vector.JPG]] <br /> <br /> Date:&amp;nbsp;August 17, 2012&lt;br&gt;<br /> <br /> Digested pDB.His.PfuMBP 1129 with Nde1 + BamH1 (see lane 1)&lt;br&gt;<br /> <br /> [[Image:1217_Vector_Digestion.JPG]]<br /> <br /> === Colony Screening ===<br /> <br /> [[Image:1217 screening.JPG]]&lt;br&gt; <br /> <br /> Digested with RE Pst + Xba1&lt;br&gt; <br /> <br /> Very light band at expected 1383 bp, very unclear&lt;br&gt; <br /> <br /> Sequencing result is not good, need to do it again.&lt;br&gt; <br /> <br /> Date:&amp;nbsp;August 20, 2012 <br /> <br /> [[Image:1217 0820 Screening.JPG]] <br /> <br /> <br /> <br /> Digest with Pst + Xba1---&amp;gt;get a band of 1272 bp&lt;br&gt;Sent sample 5 to sequencing &lt;br&gt;<br /> <br /> Clones 1, 2, 3, 5 may be correct<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