https://eln.excellgen.com/index.php?action=history&feed=atom&title=1134_pTXB1-Ecoli-Ligase 1134 pTXB1-Ecoli-Ligase - Revision history 2026-10-18T05:16:51Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1134_pTXB1-Ecoli-Ligase&diff=5467&oldid=prev Xiaojing.Wang at 19:44, 6 August 2012 2012-08-06T19:44:54Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1134 pTXB1-Ecoli-Ligase&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1134 pTXB1-Ecoli-Ligase<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;1134 <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 24, 2012&lt;br&gt; <br /> <br /> [[Image:0724 Amp.JPG]]&lt;br&gt; <br /> <br /> See lane 3, expected size 2017 bp&lt;br&gt; <br /> <br /> PCR using these primers and genomic DNA fromm MG 1665 &lt;br&gt;Ecoli_lig-Nde1-F&lt;br&gt;atcatcCATatggaatcaatcgaacaacaac <br /> <br /> Ecoli_lig-Xho1-R <br /> <br /> (gaaatgctgcgtttgctgggtagc CTCGAG catgca)&lt;br&gt;tgcatgCTCGAGgctacccagcaaacgcagcatttc <br /> <br /> Clone into pTXB1-PT-ELK (1254)&amp;nbsp;with RE&amp;nbsp;(Nde1 + Xho1).&lt;br&gt; <br /> <br /> [[Image:0726 Digestion.JPG]] <br /> <br /> See lane 8, digested insert and vector together in the same lane. Insert is missing, need to reamplify. Save vector <br /> <br /> [[Image:0724 Amp 2.JPG]]&lt;br&gt; <br /> <br /> See lane 2, expected size 2017 bp <br /> <br /> PCR using these primers and genomic DNA from MG 1665&lt;br&gt;Ecoli_lig-Nde1-F&lt;br&gt;atcatcCATatggaatcaatcgaacaacaac <br /> <br /> Ecoli_lig-Xho1-R <br /> <br /> (gaaatgctgcgtttgctgggtagc CTCGAG catgca)&lt;br&gt;tgcatgCTCGAGgctacccagcaaacgcagcatttc <br /> <br /> Clone into pTXB1-PT-ELK (1254) with RE (Nde1 + Xho1).&lt;br&gt; <br /> <br /> [[Image:0724 Digestion.JPG]]&lt;br&gt; <br /> <br /> Digested with Nde1 and Xho1 (see lane 2), cut out band, purify and ligate together with vector saved form earlier. Ligate over night, transform tomorrow morning&lt;br&gt; <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 3, expected size 2017 bp <br /> <br /> PCR using these primers and genomic DNA fromm MG 1665&lt;br&gt;Ecoli_lig-Nde1-F&lt;br&gt;atcatcCATatggaatcaatcgaacaacaac <br /> <br /> Ecoli_lig-Xho1-R <br /> <br /> (gaaatgctgcgtttgctgggtagc CTCGAG catgca)&lt;br&gt;tgcatgCTCGAGgctacccagcaaacgcagcatttc <br /> <br /> Clone into pTXB1-PT-ELK (1254) with RE (Nde1 + Xho1).&lt;br&gt; <br /> <br /> [[Image:1134_Digestion.JPG]]<br /> <br /> Ligate together, transform<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, expected size 2017 bp <br /> <br /> See lanes 19- 24 <br /> <br /> Lane 21 (sample 3)&amp;nbsp;may have insert?&amp;nbsp;Send sample 3 to sequencing <br /> <br /> Date:&amp;nbsp;August 6, 2012&lt;br&gt;<br /> <br /> [[Image:1134_0806_Screening.JPG]]&lt;br&gt;<br /> <br /> Digested MiniPrep with RE Nde1 and Xho1, expected size 2017 bp &lt;br&gt;<br /> <br /> No inserts<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> Xiaojing.Wang