https://eln.excellgen.com/index.php?action=history&feed=atom&title=2166_pT7A3E-T4-endo-VII 2166 pT7A3E-T4-endo-VII - Revision history 2026-10-18T16:06:41Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=2166_pT7A3E-T4-endo-VII&diff=13081&oldid=prev Excellgen at 18:30, 12 September 2014 2014-09-12T18:30:48Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 2166 pT7A3E-T4-endo-VII&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of2166 pT7A3E-T4-endo-VII&#039;&#039;&#039;==&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2014-8---- <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:2166&#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 Genomic DNA <br /> <br /> 1st fragment <br /> <br /> T4-endo-VII-Nde1-F atcga catatgttattgactggcaaattatac <br /> <br /> T4-endo-VII-R1 caacaccttgtccctttaagccagaacg <br /> <br /> 2nd fragment <br /> <br /> T4-endo-VII-F1cgttctggcttaaagggacaaggtgttg <br /> <br /> T4-endo-VII-Xho1-R2 (cttagaaagagtttaaaa ctcgag ctagca) tgctagctcgagttttaaactctttctaag <br /> <br /> [[Image:2116 2240 2241pcr.JPG]] <br /> <br /> join two fragments <br /> <br /> T4-endo-VII-Nde1-F atcga catatgttattgactggcaaattatac <br /> <br /> T4-endo-VII-Xho1-R2 (cttagaaagagtttaaaa ctcgag ctagca) tgctagctcgagttttaaactctttctaag <br /> <br /> [[Image:2166 1319-1pcr.JPG]]&lt;br&gt; clone into 1892 using Nde1 +Xho1&lt;br&gt; <br /> <br /> [[Image:Digest for2012fix 2166.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2014_7 <br /> <br /> Make miniprep, use Nde1 +Xho1 to find positive clone&lt;br&gt; <br /> <br /> [[Image:2012FIX 2303 1578 2166digest.JPG]][[Image:2166digest.JPG]] <br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Cloning of 2166 pT7A3E-T4-endo-VII&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of2166 pT7A3E-T4-endo-VII&#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:2267&#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 1562<br /> <br /> [[Image:2166 2302 2267PCR.JPG]]&lt;br&gt; digest with Nde1 + Xho1&lt;br&gt; <br /> <br /> [[Image:DIGEST FOR 2166 2302 2267 2336PCR.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt;&lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2014_8&lt;br&gt; <br /> <br /> Make miniprep, use&amp;nbsp; Nde1 + Xho1to find positive clone&lt;br&gt; <br /> <br /> [[Image:2166 2267 2336DIGEST.JPG]]&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 /> == &#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;&lt;br&gt; <br /> <br /> [[Category:Cloning]]</div> Excellgen