https://eln.excellgen.com/index.php?action=history&feed=atom&title=1141_p8H-DNA-Pol-1 1141 p8H-DNA-Pol-1 - Revision history 2026-10-18T06:32:03Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1141_p8H-DNA-Pol-1&diff=7290&oldid=prev Paul.xu at 18:47, 3 December 2012 2012-12-03T18:47:53Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1141 p8H-DNA-Pol-1&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1141 p8H-DNA-Pol-1 <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;1141 <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 27, 2012&lt;br&gt; <br /> <br /> [[Image:0724 Amp.JPG]]&lt;br&gt; <br /> <br /> See lane 5, expected size 2788 bp &lt;br&gt; <br /> <br /> PCR using MG1665 genomic DNA <br /> <br /> DNA-Pol1-Nde1-F&lt;br&gt;catgtaCATATGgttcagatcccccaaaatc&lt;br&gt;DNA-Pol1-Sal1-R&lt;br&gt;(cgaaaactgggatcaggcgcac GTC GAC tca cgat)&lt;br&gt;atcgtgaGTCGACGCGGCCGCCgtgcgcctgatcccagttttcg&lt;br&gt; <br /> <br /> Digest into vector 1135 with RE&amp;nbsp;Nde1 and Sal1 <br /> <br /> [[Image:0726 Digestion.JPG]]&lt;br&gt; <br /> <br /> See lane 10 (insert)&amp;nbsp;and lane 11 (vector) <br /> <br /> Cut out bands and ligate together and transform <br /> <br /> &lt;br&gt; <br /> <br /> Date:&amp;nbsp;August 2, 2012&lt;br&gt; <br /> <br /> Screening came back bad, started all over again&lt;br&gt; <br /> <br /> [[Image:0802 Amp.JPG]]&lt;br&gt; <br /> <br /> See lane 4, expected size 2788 bp <br /> <br /> PCR using MG1665 genomic DNA <br /> <br /> DNA-Pol1-Nde1-F&lt;br&gt;catgtaCATATGgttcagatcccccaaaatc&lt;br&gt;DNA-Pol1-Sal1-R&lt;br&gt;(cgaaaactgggatcaggcgcac GTC GAC tca cgat)&lt;br&gt;atcgtgaGTCGACGCGGCCGCCgtgcgcctgatcccagttttcg <br /> <br /> 1[[Image:1141 Digestion.JPG]] <br /> <br /> Ligate together, transform <br /> <br /> Digest into vector 1135 with RE Nde1 and Sal1 &lt;br&gt; <br /> <br /> === &lt;u&gt;&lt;/u&gt;Colony Screening&lt;br&gt; ===<br /> <br /> Date: July 27, 2012 <br /> <br /> [[Image:0727 Digestion 2.JPG]] <br /> <br /> Digested MiniPrep with RE Nde1 and Sal1, expected size 2788 bp <br /> <br /> Samples 2, 5 and 6 may be correct, submitted sample 5 for sequencing&lt;br&gt; <br /> <br /> Date:&amp;nbsp;August 6, 2012&lt;br&gt; <br /> <br /> 1[[Image:1141 0806 Screening.JPG]]&lt;br&gt; <br /> <br /> Digested MiniPrep with RE Nde1 and Sal1, expected size 2788 bp &lt;br&gt; <br /> <br /> No inserts <br /> <br /> &lt;br&gt; <br /> <br /> Date: Nov30, 2012 &lt;br&gt; <br /> <br /> [[Image:1141 1147 1277 1444PCR.JPG]]&lt;br&gt; <br /> <br /> Digest vector 1135 and 1141pcr products with Nde1 and Sal1 &lt;br&gt; <br /> <br /> [[Image:1141PCR 1135 1147PCR 1135DIGEST.JPG]] <br /> <br /> &lt;br&gt; Digested MiniPrep with RE Nde1 and Sal1, expected size 2788 bp <br /> <br /> [[Image:1141digest.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