https://eln.excellgen.com/index.php?action=history&feed=atom&title=1117_p8H-Klenow-Exo- 1117 p8H-Klenow-Exo- - Revision history 2026-10-18T02:19:59Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1117_p8H-Klenow-Exo-&diff=7439&oldid=prev Paul.xu at 15:25, 12 December 2012 2012-12-12T15:25:57Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1117 p8H-Klenow-Exo-&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1117 p8H-Klenow-Exo- <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;July 23, 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;1117 <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 23, 2012&lt;br&gt; <br /> <br /> [[Image:1117 Amp.JPG]]&lt;br&gt; <br /> <br /> PCR using MG1665 genomic DNA <br /> <br /> KlenowExo-_Nde1-F <br /> <br /> cagtcg catatg ATTGAACGTAACGGTGTGAAG <br /> <br /> KlenowExo-_Xho1-R&lt;br&gt;(GAAAACTGGGATCAGGCGCAC CTCGAG ctgtca)&lt;br&gt;tgacagCTCGAGGTGCGCCTGATCCCAGTTTTC&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> Expected size 1168 bp <br /> <br /> Cut out bands in lanes 1 and 2, purify and digest with RE&amp;nbsp;Nde1 and Xho1 with vector p8H <br /> <br /> [[Image:0726 Digestion.JPG]] <br /> <br /> See lanes 2 and 3, cut out both bands and ligate together and transform <br /> <br /> Date: August 2, 2012&lt;br&gt; <br /> <br /> Sequencing came back bad, start over again&lt;br&gt; <br /> <br /> [[Image:0802 Amp.JPG]]&lt;br&gt; <br /> <br /> PCR using MG1665 genomic DNA <br /> <br /> KlenowExo-_Nde1-F <br /> <br /> cagtcg catatg ATTGAACGTAACGGTGTGAAG <br /> <br /> KlenowExo-_Xho1-R&lt;br&gt;(GAAAACTGGGATCAGGCGCAC CTCGAG ctgtca)&lt;br&gt;tgacagCTCGAGGTGCGCCTGATCCCAGTTTTC <br /> <br /> &lt;br&gt;Expected size 1168 bp <br /> <br /> Cut out bands in lanes 1, purify and digest with RE Nde1 and Xho1 with vector p8H &lt;br&gt; <br /> <br /> 1[[Image:1117 Digestion.JPG]]&lt;br&gt; <br /> <br /> See lanes 1 and 2, cut out both bands and 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;July 27, 2012 <br /> <br /> [[Image:0727 Digestion.JPG]] <br /> <br /> Digested MiniPrep with RE Nde1 and Xho1. See lanes 1-6, expected size ~1160 bp. Samples 2 and 4 may be correc, send sample 2 to sequencing.&amp;nbsp; <br /> <br /> Date:&amp;nbsp;August 6, 2012&lt;br&gt; <br /> <br /> [[Image:1117 0806 Screening.JPG]]&lt;br&gt; <br /> <br /> Digested MiniPrep with RE Nde1 and Xho1. See lanes 1-6, expected size ~1160 bp&lt;br&gt; <br /> <br /> Lanes 1, 2, 3, 4 may be correct <br /> <br /> Send sample 2 to sequencing <br /> <br /> &lt;br&gt; <br /> <br /> Purpose: Cloning of 1117 p8H-Klenow-Exo- <br /> <br /> Date: Dec, 2012 <br /> <br /> Notebook: <br /> <br /> Page: <br /> <br /> Clone ID: 1117 <br /> <br /> Experimenters: Shanyi&lt;br&gt; <br /> <br /> PCR&lt;br&gt; <br /> <br /> [[Image:1138 1135digest 1277 1117 1118pcr.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> DIGEST 1422 and 1277 pcr products with Nde1 + Xho1&lt;br&gt; <br /> <br /> [[Image:1117PCR-1118PCR 1422 1277 V15 1292 369 1139 1141 1147.JPG.JPG]] <br /> <br /> SCREENING<br /> [[Image:1481 screening 2012 12 7.jpg]]&lt;br&gt; <br /> <br /> ==<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