https://eln.excellgen.com/index.php?action=history&feed=atom&title=2405-pEC-Miraculin-Term2405-pEC-Miraculin-Term - Revision history2026-10-18T20:59:12ZRevision history for this page on the wikiMediaWiki 1.44.0https://eln.excellgen.com/index.php?title=2405-pEC-Miraculin-Term&diff=14007&oldid=prevExcellgen at 20:31, 13 March 20152015-03-13T20:31:20Z<p></p>
<p><b>New page</b></p><div>== '''Cloning of 2412 pEC2-AaeMutS''' ==<br />
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'''Purpose:''' Cloning of 2412 pEC2-AaeMutS<br />
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'''Date:&nbsp;''' 2015-3-5<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:''' Paul & Xu Xin<br />
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== '''Results''' ==<br />
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=== cloning ===<br />
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1.Use plant as template to extract genomic DNA.<br />
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2. PCR use the genomic DNA as template.<br />
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&nbsp;&nbsp;&nbsp; The 1st: the DNA extacted by Paul; the 2nd PCR: The DNA extacted by Xin<br />
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[[File:2405 3 6-2015- PCR.JPG|RTENOTITLE]]<br />
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3. Gel purified the PCR#2,<br />
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4. Digestion, clean up digestion (with NcoI/KpnI), ligation. transformaiton.<br />
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5. select 10 colonies from transformation plate to PCR screening.<br />
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[[File:2405-PCR-screening transformation-3-11-2015.JPG|RTENOTITLE]]<br />
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6. Select colonies 1,2,7,8,9 and 10 to innoculate.<br />
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7. extact plasmid from the step 6.<br />
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8. Digestion the plasmids with NcoI/KpnI<br />
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[[File:2405 confirmation digestion 3 11 2015.JPG|RTENOTITLE]]<br />
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9. From the gel, no insection in these 6 plasmid.<br />
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10. As Paul curious what is the product,PCR the these 6 selected plasmids. Also PCR the genmoic DNA as step 1.<br />
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== '''[[File:2405 3 13-2015-transformation plasmid PCR.JPG 400 × 400, 46 KB|RTENOTITLE]]''' ==<br />
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Send plasmid NO. 7 to sequence.<br />
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Also the genomic DNA PCR was purified, but elution in 40ul (instead of 20) and casue to low concentration ( be careful for the next step), maybe need to rewash and elution in a low concentration.<br />
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Date:&nbsp;<br />
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== '''Result intepretation/Summary''' ==<br />
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== '''Conclusion''' ==<br />
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== '''&nbsp;''' ==<br />
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== '''&nbsp;''' ==<br />
[[Category:Cloning]]</div>Excellgen