https://eln.excellgen.com/index.php?action=history&feed=atom&title=2407_pCAMBIA-Miraculin2407 pCAMBIA-Miraculin - Revision history2026-10-18T11:52:07ZRevision history for this page on the wikiMediaWiki 1.44.0https://eln.excellgen.com/index.php?title=2407_pCAMBIA-Miraculin&diff=14114&oldid=prevExcellgen at 17:20, 1 April 20152015-04-01T17:20:44Z<p></p>
<p><b>New page</b></p><div>== '''Cloning of 2407''' ==<br />
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'''Purpose:''' Cloning of 2407 pCAMBIA-35s promoter-Miraculin-terminator<br />
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'''Date:&nbsp;''' 2015-3-24<br />
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'''Notebook:&nbsp;'''<br />
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'''Page:&nbsp;'''<br />
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'''Clone ID: 2412'''<br />
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'''Experimenters:''' Xin<br />
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subclone 2422 promoter-miraculin-terminator into vector 1153.<br />
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&nbsp;[[File:2407-digest.jPG|RTENOTITLE]]<br />
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== ==<br />
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ligation, transformation<br />
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PCR screening: as the PCR cycle is 25, so no strong band can be found on on the gel photo.&nbsp;<br />
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But there is a band on the gel under dark for all the colony on 400 bp location. Select the colony 2,4,and 5 mini plasmid prepation.<br />
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== '''[[File:2407-screening-3-27-15.jPG|RTENOTITLE]]''' ==<br />
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After analysis, found out the selection of insert was wrong. It should be1600bp instead of 700bp.<br />
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Paul redo it again.<br />
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[[File:2407 digestion paul 3-27-15.JPG|RTENOTITLE]]<br />
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Select 8 colonies for screening.<br />
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[[File:2407 digestion 3-30-15.JPG|RTENOTITLE]]<br />
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'''&nbsp;'''<br />
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&nbsp;[[File:2407 redigest 3-30-15.JPG|RTENOTITLE]]<br />
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Reselect some colonies to screen, by PCR the primers are:<br />
<pre>2405-Miraculin-Term-Kpn1-F </pre><br />
<pre>pCAMBIA-2301-Nos-seq-R</pre><br />
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[[File:2407 screening 3-31-15.JPG|RTENOTITLE]] The colony # 4, 5, 6,10,11,12 16 have the right size product -600bp.<br />
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[[File:2407_screening_4-1.JPG]]<br />
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== '''Conclusion''' ==<br />
[[Category:Cloning]]</div>Excellgen