https://eln.excellgen.com/index.php?action=history&feed=atom&title=1277_pET30b-RNaseIII 1277 pET30b-RNaseIII - Revision history 2026-10-18T05:50:22Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1277_pET30b-RNaseIII&diff=7444&oldid=prev Paul.xu at 15:29, 12 December 2012 2012-12-12T15:29:58Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1277 pET30b-RNaseIII&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1277 pET30b-RNaseIII<br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;July 30, 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;1277 <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 /> Date:&amp;nbsp;July 30, 2012&lt;br&gt; <br /> <br /> [[Image:0730 Amp.JPG]]&lt;br&gt; <br /> <br /> See lane 4&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> PCR using genomic DNA from K-12 substr. MG1655&lt;br&gt;RnaseIII-Nde1-F AGCTCTCATatgaaccccatcgtaattaatc&lt;br&gt;RnaseIII-Xho1-R gtctcgctcgagttccagctccagttttttcaac&lt;br&gt;clone into Nde1 + Xho1 digested 1254 pTXB1-PT-ELK <br /> <br /> Expected size 1311 bp&lt;br&gt; <br /> <br /> Cut out band and save in box <br /> <br /> Date: August 1, 2012&lt;br&gt; <br /> <br /> [[Image:0801 Digestion.JPG]]136&lt;br&gt; <br /> <br /> Digested insert (lane 4)&amp;nbsp;with RE Nde1 + Xho1, vector is not good, digest again tomorrow, join with lane 4, purify and ligate together &lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date:&amp;nbsp;August 6, 2012 <br /> <br /> Digested MiniPrep with RE Xho1 and Nde1, expected size 1311 bp <br /> <br /> [[Image:1277 0806 Screening.JPG]]136 <br /> <br /> All negative for inserts <br /> <br /> PCR&lt;br&gt;<br /> <br /> [[Image:1138_1135digest_1277_1117_1118pcr.JPG]]&lt;br&gt;<br /> <br /> DIGEST V15pET30b and 1277pcr produts with Nde1 + Xho1&lt;br&gt;<br /> <br /> [[Image:1117PCR-1118PCR_1422_1277_V15_1292_369_1139_1141_1147.JPG.JPG]]<br /> <br /> <br /> SCREENING<br /> [[Image:1481 screening 2012 12 7.jpg]]&lt;br&gt; <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