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760 pEC2-Pme1 - Revision history
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Excellgen at 17:31, 15 April 2015
2015-04-15T17:31:58Z
<p></p>
<p><b>New page</b></p><div>== '''Cloning of760 pEC2-Pme1''' ==<br />
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'''Purpose:''' Cloning of760 pEC2-Pme1'''=='''<br />
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'''Date: '''2014-12_2015_1<br />
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'''Notebook:0'''<br />
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'''Page:'''<br />
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'''Clone ID:760'''<br />
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'''Experimenters:'''&nbsp; shanyi<br />
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== '''Results''' ==<br />
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=== CLONING ===<br />
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gene synthesis<br />
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[[File:760 761 2391fixpcr.JPG|RTENOTITLE]]<br />
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clone to 2015 using Nde1 + Xho1<br />
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[[File:Digest for 760 761.JPG|RTENOTITLE]]<br />
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=== Colony Screening ===<br />
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Date: 2015_1<br />
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Make miniprep, use Nde1 + Xho1 to find positive clone<br />
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<br/>[[File:2391fix 758 760 761 757 )2392digest.JPG|RTENOTITLE]]<br />
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=== Fix 760-5 ===<br />
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pcr<br />
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fragment 1<br />
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761-PmeI-1+761-PmeI-14<br />
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fragment 2 761-PmeI-15+761-PmeI-18<br />
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[[File:758fix 760fix 2403 2401-2pcr.JPG|RTENOTITLE]]<br />
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clone to 2015 using Nde1 + Xho1<br />
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[[File:758fix 760fix 2401 2321 2399pcr.JPG|RTENOTITLE]]<br />
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=== Colony Screening ===<br />
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Date: 2015_1<br />
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Make miniprep, use Nde1 + Xho1 to find positive clone<br />
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[[File:760fix 2399digest.JPG|RTENOTITLE]]<br />
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=== Fix 760-1 ===<br />
<br />
pcr<br />
<br />
[[File:760FIX2PCR.JPG|RTENOTITLE]]<br />
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clone to 2015 using Nde1 + Xho1<br />
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[[File:DIGEST FOR 760 2387 2409 2410.JPG|RTENOTITLE]]<br />
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=== Colony Screening ===<br />
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Date: 2015_1<br />
<br />
Make miniprep, use Nde1 + BamH1 to find positive clone<br />
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[[File:760n 2387n 2387digest.JPG|RTENOTITLE]]<br />
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== '''760-fix mutation''' ==<br />
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Missing CT<br />
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Pick up 760-fix-5 as template<br />
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PCR 3-26-15<br />
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[[File:760-FIX-3-26-15.JPG|RTENOTITLE]]<br />
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3-27-15 Repeat of 3-26<br />
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[[File:760-fix-0327.png|RTENOTITLE]]<br />
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4-8-15 760 insert PCR product<br />
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[[File:760 insert PCR.JPG|RTENOTITLE]]<br />
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== 760 clone ==<br />
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2015-4-14<br />
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[[File:760 clones-1.JPG|RTENOTITLE]]<br />
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[[File:760 clones-2.jpg|RTENOTITLE]]<br />
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== 760 PCR procuct digestion after innoculation ==<br />
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[[File:760 PCR product digestion.JPG]]<br />
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== '''Result intepretation/Summary''' ==<br />
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*<br />
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== '''Conclusion''' ==<br />
[[Category:Cloning]]</div>
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