https://eln.excellgen.com/index.php?action=history&feed=atom&title=2061_pEC-ExcellScript-II 2061 pEC-ExcellScript-II - Revision history 2026-10-18T15:39:06Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=2061_pEC-ExcellScript-II&diff=11190&oldid=prev Paul.xu: Created page with "== '''Cloning of 2061 pEC-ExcellScript-II''' == '''Purpose:''' Cloning of2061 pEC-ExcellScript-II'''==''' '''Date: '''2014-2---- '''Notebook:0''' '''Page:''' '''Clone I..." 2014-02-07T21:30:58Z <p>Created page with &quot;== &#039;&#039;&#039;Cloning of 2061 pEC-ExcellScript-II&#039;&#039;&#039; == &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of2061 pEC-ExcellScript-II&#039;&#039;&#039;==&#039;&#039;&#039; &#039;&#039;&#039;Date: &#039;&#039;&#039;2014-2---- &#039;&#039;&#039;Notebook:0&#039;&#039;&#039; &#039;&#039;&#039;Page:&#039;&#039;&#039; &#039;&#039;&#039;Clone I...&quot;</p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 2061 pEC-ExcellScript-II&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of2061 pEC-ExcellScript-II&#039;&#039;&#039;==&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2014-2---- <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:2061&#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 <br /> <br /> PCR two fragments:&lt;br&gt;Fragment 1&lt;br&gt;ssIII-Nde1-F1&lt;br&gt;ctagcCATATGACCCTAAATATAGAAGATGAGCATCGGCTAC<br /> <br /> ssIII-M289L-R4&lt;br&gt;(GAGGCCAGAAAA GAGACTGTG cTG GGG CAGCCTACTCCGAAG ACCCCTCGACAACTAAG)&lt;br&gt;CTTAGTTGTCGAGGGGTCTTCGGAGTAGGCTGCCCCAgCACAGTCTCTTTTCTGGCCTC<br /> <br /> Fragment 2&lt;br&gt;ssIII-V223H-F4&lt;br&gt;CAGACTTCCGGATCCAGCACCCAGACTTGATCCTGCTACAGTACcatGATGACTTACTGCTG<br /> <br /> ssIII-Xho1-R8&lt;br&gt;(TCACAGAGAATCCAGACACCTCTACCCTCCTCATAGAAAATTCATCACCGAATTCCCGCTTAATTAATCTCGAGctag)&lt;br&gt;ctagCTCGAGATTAATTAAGCGGGAATTCGGTGATGAATTTTCTATGAGGAGGGTAGAGGTGTCTGGATTCTCTGTGA&lt;br&gt;<br /> <br /> &lt;br&gt;<br /> <br /> [[Image:2061firstpcr.JPG]]&lt;br&gt;2. Join two fragments using&lt;br&gt;ssIII-Nde1-F1 + ssIII-Xho1-R8&lt;br&gt;&lt;br&gt;<br /> <br /> [[Image:2061pcr.JPG]]&lt;br&gt;<br /> <br /> 3. Digest with Nde1 + Xho1, clone into pEC&lt;br&gt;&lt;br&gt;&lt;br&gt;<br /> <br /> &lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2013-10-23 <br /> <br /> Make miniprep, digest with Nde1 + Xho1+BamHi&lt;br&gt; <br /> <br /> [[Image:2028_2038_2059_2061DIGEST.JPG]]<br /> <br /> &lt;br&gt; <br /> <br /> &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 /> [[Category:Cloning]]</div> Paul.xu