https://eln.excellgen.com/index.php?action=history&feed=atom&title=1341_pDB.His.MBP-%283C%29-p53wt 1341 pDB.His.MBP-(3C)-p53wt - Revision history 2026-10-18T05:51:15Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1341_pDB.His.MBP-(3C)-p53wt&diff=6044&oldid=prev Paul.xu at 16:00, 14 September 2012 2012-09-14T16:00:53Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1341 pDB.His.MBP-(3C)-p53wt&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1341 pDB.His.MBP-(3C)-p53wt <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;August 24, 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;1341 <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 24, 2012 <br /> <br /> PCR from 80 pLex-p53wt <br /> <br /> P53-Nhe1-F&lt;br&gt;atcgac GCTAGC ATGGAGGAGCCGCAGTCAGATC <br /> <br /> P53-EcoR1-R1&lt;br&gt;(GAAGGGCCTGACTCAGACTGA gaattc atcgac)&lt;br&gt;gtcgatgaattcTCAGTCTGAGTCAGGCCCTTC&lt;br&gt; <br /> <br /> See lane 3 <br /> <br /> [[Image:1341 Amp.JPG]] <br /> <br /> Digested DNA (lane 3)&amp;nbsp;with RE&amp;nbsp;Nhe1and EcoR1 <br /> <br /> [[Image:0824 Digestion.JPG]] <br /> <br /> Digested vector 1207 with RE&amp;nbsp;Nhe1 and EcoR1 (see lane 2) <br /> <br /> [[Image:Vector Digestions.JPG]] <br /> <br /> Cut out bands, purifiy and ligate together, transform <br /> <br /> === &lt;u&gt;&lt;/u&gt;Colony Screening&lt;br&gt; ===<br /> <br /> &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 /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Cloning of 1341 pDB.His.MBP-(3C)-p53wt&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1341 pDB.His.MBP-(3C)-p53wt <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;August 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;1341 <br /> <br /> &#039;&#039;&#039;Experimenters: &#039;&#039;&#039;yingying lu <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === Cloning&lt;br&gt; ===<br /> <br /> Date:&amp;nbsp;August 30, 2012 <br /> <br /> PCR from 80 pLex-p53wt <br /> <br /> P53-Nhe1-F&lt;br&gt;atcgac GCTAGC ATGGAGGAGCCGCAGTCAGATC <br /> <br /> P53-EcoR1-R1&lt;br&gt;(GAAGGGCCTGACTCAGACTGA gaattc atcgac)&lt;br&gt;gtcgatgaattcTCAGTCTGAGTCAGGCCCTTC&lt;br&gt; <br /> <br /> [[Image:1331+1341+1352 PCR 8-30-2012.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> Digested DNA (lane )&amp;nbsp;with RE&amp;nbsp;Nhe1and EcoR1 <br /> <br /> &lt;br&gt; <br /> <br /> Digested vector 1207(lane4) with&amp;nbsp; Nhe1 and EcoR1&amp;nbsp; <br /> <br /> [[Image:1341+1331+1352 clone 8-30-2012.JPG]] <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> Cut out bands, purifiy and ligate together, transform <br /> <br /> === &lt;u&gt;&lt;/u&gt;Colony Screening&lt;br&gt; ===<br /> <br /> 2012-9-5 <br /> <br /> [[Image:1256+1341 digest detct 9-5-2012.JPG]] <br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> sequence result: not the right colony&lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Cloning of 1341 pDB.His.MBP-(3C)-p53wt&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1341 pDB.His.MBP-(3C)-p53wt <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2012-9-12---2012-9-14&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;1341 <br /> <br /> &#039;&#039;&#039;Experimenters: &#039;&#039;&#039;yingying lu <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === Cloning&lt;br&gt; ===<br /> <br /> Date:&amp;nbsp;August 30, 2012 <br /> <br /> PCR from 80 pLex-p53wt <br /> <br /> P53-Nhe1-F&lt;br&gt;atcgac GCTAGC ATGGAGGAGCCGCAGTCAGATC <br /> <br /> P53-EcoR1-R1&lt;br&gt;(GAAGGGCCTGACTCAGACTGA gaattc atcgac)&lt;br&gt;gtcgatgaattcTCAGTCTGAGTCAGGCCCTTC&lt;br&gt; <br /> <br /> [[Image:1331+1341+1352 PCR 8-30-2012.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> Digested DNA (lane )&amp;nbsp;with RE&amp;nbsp;Nhe1and EcoR1 <br /> <br /> &lt;br&gt; <br /> <br /> Digested vector 1342 (lane4) with&amp;nbsp; Nhe1 and EcoR1&amp;nbsp; <br /> <br /> [[Image:1289+1341+1346+1348-2 clone 9-12-2012.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> Cut out bands, purifiy and ligate together, transform <br /> <br /> === &lt;u&gt;&lt;/u&gt;Colony Screening&lt;br&gt; ===<br /> <br /> 2012-9-14 <br /> <br /> mini prep and digest :Nhe1+EcoR1&lt;br&gt;<br /> <br /> [[Image:1341+1348_digest_detect_9-14-2012.JPG]]<br /> <br /> <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 /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> &lt;br&gt; <br /> <br /> [[Category:Cloning]]</div> Paul.xu