https://eln.excellgen.com/index.php?action=history&feed=atom&title=27952795 - Revision history2026-10-18T09:24:50ZRevision history for this page on the wikiMediaWiki 1.44.0https://eln.excellgen.com/index.php?title=2795&diff=21018&oldid=prevPaul.xu at 16:45, 19 June 20202020-06-19T16:45:20Z<p></p>
<p><b>New page</b></p><div>== 2795 gibson ==<br />
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[[File:2795 Gibson 1.JPG|RTENOTITLE]]<br />
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[[File:2746 2795 gibson.JPG|RTENOTITLE]]<br />
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[[File:2795 gibson vector.JPG|RTENOTITLE]]<br />
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== 2795 subclone ==<br />
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20200617 digest 2795_pJet2.1 with NdeI/NotI. The size should be 2181bp, but sometimes the maker is not that accurate. So we select two bands for further ligation with the vector 2794 digested with&nbsp;NdeI/NotI.<br />
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[[File:2746 2773 2779 2795 pJet digestion.JPG|RTENOTITLE]]<br />
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20200618 2795 vector comes from 2794 digested with&nbsp;NdeI/NotI. Time course showed the two hours digestion looks best.&nbsp;<br />
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[[File:2795 subclone vector NdeI NcoI.JPG|RTENOTITLE]]<br />
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20200619 2795-T7-VEE clone PCR was performed. Primers used will produce 2153bp product. So the insert band between 3-5kb looks like the one we want.&nbsp;<br />
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[[File:2746 2795 clone PCR.JPG|RTENOTITLE]]<br />
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== PCR ==<br />
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== 2795 mRNA cleancap ==<br />
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[[File:2795 1st PCR.JPG|RTENOTITLE]]<br />
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[[File:2795 2nd PCR.JPG|RTENOTITLE]]<br />
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[[File:2795 PCR product confirmation.JPG|RTENOTITLE]]<br />
[[Category:Cloning]]</div>Paul.xu