https://eln.excellgen.com/index.php?action=history&feed=atom&title=2240_pEC2-T4-uvsX 2240 pEC2-T4-uvsX - Revision history 2026-10-18T08:34:49Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=2240_pEC2-T4-uvsX&diff=13753&oldid=prev Excellgen at 17:58, 26 December 2014 2014-12-26T17:58:46Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 2240 pEc2-T4-uvsX&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of2240 pEc2-T4-uvsX-&#039;&#039;&#039;==&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2014-9---- <br /> <br /> &#039;&#039;&#039;Notebook:0&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID:2240&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039;&amp;nbsp; shanyi <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === CLONING ===<br /> <br /> PCR from T4 phage Genomic DNA &lt;br&gt; <br /> <br /> Fragment 1&lt;br&gt; <br /> <br /> T4-uvsX-Nde1-F tcgaacatatgtctgatttaaaatctcgtttg &lt;br&gt; <br /> <br /> T4-uvsX-R1 gtgtttcgtatgtatggttaatagc <br /> <br /> [[Image:2116 2240 2241pcr.JPG]]&lt;br&gt; <br /> <br /> Fragment 2 &lt;br&gt; <br /> <br /> Cloning Method <br /> <br /> gene synthesis&lt;br&gt; <br /> <br /> [[Image:2166ns-1319 2240 1504 2241pcr.JPG]]&lt;br&gt; <br /> <br /> Join two fragments <br /> <br /> T4-uvsX-Nde1-F tcgaacatatgtctgatttaaaatctcgtttg &lt;br&gt; <br /> <br /> T4-uvsX-Xho1-R (gatatggaagaatttaatgaa ctcgag tagca) tgctactcgagttcattaaattcttccatatc <br /> <br /> digest with Nde1 + Xho1&lt;br&gt; <br /> <br /> [[Image:DIGEST FOR 2297 1504 1319 2240 2241 2246 2199.JPG]] <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2014_8&lt;br&gt; <br /> <br /> Make miniprep, use&amp;nbsp; Nde1 + Xho1 to find positive clone&lt;br&gt; <br /> <br /> [[Image:1319A 1504A 2199A 2240A 2241A 2297DIGEST.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> sequence 2240a-5 with M13F primer, the second fragment is right.&lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Cloning of 2240 pEc2-T4-uvsX&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of2240 pEc2-T4-uvsX-&#039;&#039;&#039;==&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2014-9---- <br /> <br /> &#039;&#039;&#039;Notebook:0&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID:2240&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039;&amp;nbsp; shanyi <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === CLONING ===<br /> <br /> gene synthesis the 1st fragment,pcr the 2nd fragment from 2240a-5 <br /> <br /> [[Image:2350 1917 2240pcr.JPG]]&lt;br&gt; jOIN two fragment by PCR <br /> <br /> [[Image:736fix 1916fix2 1917 2240.JPG]]&lt;br&gt; digest with Nde1 + XhoI&lt;br&gt; <br /> <br /> [[Image:Digest for2354 2369 736fix2 1916fix3 2240.JPG]][[Image:Digest for736fix2 1916fix3 2240.JPG]]&lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2014_8&lt;br&gt; <br /> <br /> Make miniprep, use&amp;nbsp; Nde1 + Xho1 to find positive clone&lt;br&gt; <br /> <br /> [[Image:736FIX2 1916FIX3 2240 2354 2369 2375.JPG]]&lt;br&gt; <br /> <br /> [[Image:736n_1916n_1917n_2240n_2352_2353digest.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 /> [[Category:Cloning]]</div> Excellgen