https://eln.excellgen.com/index.php?action=history&feed=atom&title=2436-pET30-30GFP 2026-10-18T11:23:53Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=2436-pET30-30GFP&diff=14324&oldid=prev 2015-05-04T20:23:57Z

<p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of &lt;span dir=&quot;auto&quot;&gt;2436-pET30-30GFP&lt;/span&gt;&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Date:&amp;nbsp;&#039;&#039;&#039; 2015-April<br /> <br /> &#039;&#039;&#039;&#039;&#039;Clone ID&#039;&#039;&#039;: 2436&#039;&#039;<br /> <br /> &#039;&#039;&#039;Experiment&#039;&#039;&#039;:&amp;nbsp; Xin<br /> <br /> &#039;&#039;&#039;Results&#039;&#039;&#039;:<br /> <br /> === 1.&amp;nbsp;&lt;span class=&quot;mw-headline&quot; id=&quot;Gene_Synthesis_by_PCR_using_oligo_as_template&quot;&gt;Gene Synthesis &lt;/span&gt;by two sections: ===<br /> <br /> === Section I: 1-1 __ 1-10; Section II: 1-11 __1-20 ===<br /> <br /> [[File:2423, 2436 PCR 4-9-11.JPG|319x129px|RTENOTITLE]]<br /> <br /> <br /> <br /> <br /> <br /> combine two sections together, PCR again.<br /> <br /> [[File:2423 2436 PCR 4-9-15.JPG|276x122px|RTENOTITLE]]<br /> <br /> &amp;nbsp; 2. Clone: <br /> <br /> Insert the PCR product into vector 440. <br /> <br /> [[File:440 digestion.JPG|RTENOTITLE]]<br /> <br /> Ligation and transformation.&amp;nbsp; <br /> <br /> == &#039;&#039;&#039;&#039;&#039;Screening&#039;&#039;&#039;&#039;&#039;: ==<br /> &lt;div&gt;&lt;span dir=&quot;auto&quot;&gt;&amp;nbsp;[[File:IMG 0317 2436 SCREENING1.JPG|285x162px|RTENOTITLE]]&lt;/span&gt;&amp;nbsp; The insectionthe size is about 1200bp, but should be about 850 bp.&lt;br/&gt;&lt;/div&gt;&lt;div&gt;&lt;br/&gt;&lt;/div&gt;&lt;div&gt;Redigested again using different enzyme: XhoI/ NdeI&lt;br/&gt;&lt;/div&gt;&lt;div&gt;[[File:2436-screening- II .JPG|271x186px|RTENOTITLE]]&amp;nbsp; The size is still not right.&lt;br/&gt;&lt;/div&gt;&lt;div&gt;&lt;br/&gt;&lt;/div&gt;&lt;div&gt;Selected one clony to sequence, the clone looks all right. The marker on the argrose gel isn&#039;t very accurcy.&lt;br/&gt;&lt;/div&gt;&lt;div&gt;&lt;br/&gt;&lt;/div&gt;<br /> == &#039;&#039;&#039;Fix on April, 28, 2015&#039;&#039;&#039; ==<br /> <br /> &amp;nbsp;First PCR fragment: use the oligos 2436-30-GFP-1;2436-30-GFP-12 as primers<br /> <br /> &amp;nbsp;Second PCR fragment: use the oligos 2436-30-GFP-13;2436-30-GFP-20 as primers<br /> &lt;div&gt;[[File:2423,2436,2421 PCR.JPG|337x141px|RTENOTITLE]]&lt;br/&gt;&lt;/div&gt;&lt;div&gt;&lt;br/&gt;&lt;/div&gt;&lt;div&gt;Join two PCR together:&lt;br/&gt;&lt;/div&gt;&lt;div&gt;[[File:2436, 2423, 2421 PCR.JPG|397x148px|RTENOTITLE]]&lt;br/&gt;&lt;/div&gt;&lt;div&gt;&lt;br/&gt;&lt;/div&gt;&lt;div&gt;Screening: &lt;br/&gt;&lt;/div&gt;&lt;div&gt;[[File:2434 2436 screening 5-1-15.JPG|462x148px|RTENOTITLE]]&lt;br/&gt;&lt;/div&gt;&lt;div&gt;&lt;br/&gt;&lt;/div&gt;&lt;div&gt;All 8 colonies have right insert. &lt;br/&gt;&lt;/div&gt;<br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> [[Category:Pages with broken file links]] [[Category:Cloning]]</div>

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