https://eln.excellgen.com/index.php?action=history&feed=atom&title=1916_pEC2-M-SssI 1916 pEC2-M-SssI - Revision history 2026-10-18T06:31:26Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1916_pEC2-M-SssI&diff=13530&oldid=prev Excellgen at 21:10, 31 October 2014 2014-10-31T21:10:45Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1916 pEC2-M-SssI-&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of1916 pEC2-M-SssI-&#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:1916&#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 fragment 1,Fragment 2&lt;br&gt; <br /> <br /> [[Image:1916 2326 2327 2328 2199pcr.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> join two fragments by PCR&lt;br&gt; <br /> <br /> [[Image:2328 1916 2199 2325 1504 2321 2345PCR.JPG]] <br /> <br /> digest with Nde1 + Xho1&lt;br&gt; <br /> <br /> [[Image:Digest for 1916 2199 2346.JPG]]&lt;br&gt; <br /> <br /> &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:1038 1916 2321 2297digest.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> === fix 1916-2 pEC2-M-SssI ===<br /> <br /> pcr &lt;br&gt; <br /> <br /> [[Image:2321 1319fix 1916fixpcr.JPG]]&lt;br&gt; <br /> <br /> jOIN two fragment by PCR <br /> <br /> [[Image:1916fixpcr.JPG]]&lt;br&gt; digest with Nde1 + XhoI&lt;br&gt; <br /> <br /> [[Image:Digest for 1319fix 1916fix 2360 2361 2321.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:1319FIX 1916FIX 2360 2361 2321DIGEST.JPG]]&lt;br&gt; <br /> <br /> use T7 term primer+T4-BGT-2-16 to do PCR&lt;br&gt; <br /> <br /> [[Image:1319FIX 1916FIX pcrSCREENING.JPG]] <br /> <br /> sequence 1916fix-1 not fixed,need to fix again&lt;br&gt; <br /> <br /> === fix 1916-2 pEC2-M-SssI again ===<br /> <br /> pcr &lt;br&gt; <br /> <br /> [[Image:736fix 1916fix2 2323pcr.JPG]][[Image:736fix 1916fix2 2323pcr2.JPG]] <br /> <br /> 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 /> 36_pEC2-mNco1[[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 /> &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 /> &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 /> [[Category:Cloning]]</div> Excellgen