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3009 pCMV5-Flag-H-IgG2b - Revision history
2026-10-18T19:24:45Z
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Crm at 18:30, 26 March 2018
2018-03-26T18:30:31Z
<p></p>
<p><b>New page</b></p><div>[[File:3009 3035 3010 Xho1.JPG|RTENOTITLE]]<br />
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Date: 1/24/2018<br />
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Digest miniprep DNA with Xho1<br />
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vector: pMiniT2<br />
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[[File:3001 3009 3135 950 948 digestion.JPG|RTENOTITLE]]<br />
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[[File:948-3135-3009-3001-950 digestion2.JPG.JPG|RTENOTITLE]]<br />
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1/25/2018<br />
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Miniprep DNA in pMiniT2 vector digested with Xho1<br />
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[[File:3001 3009 digestion.JPG|RTENOTITLE]]<br />
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[[File:940 3001 3009 3135 digestion2.JPG|RTENOTITLE]]<br />
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Digest of 3009pMiniT<br />
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Enzymes:BsrGI+NheI<br />
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Time:1hour<br />
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Buffer:NEB2<br />
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[[File:3009pminitedigestion1.JPG|RTENOTITLE]]<br />
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'''Digest of 3009 pMiniT'''<br />
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Enzymes:XhoI+NheI<br />
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Time:30min<br />
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Buffer:Fastfigest<br />
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[[File:3009pminit2digestion.JPG|RTENOTITLE]]<br />
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'''Subclone of 945'''<br />
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1:1727 XbaI+NcoI 173&nbsp; Cutsmart<br />
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2:1834 XbaI+NcoI 173&nbsp; Cutsmart<br />
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3:945pMiniT XbaI+NcoI 1326&nbsp; Cutsmart<br />
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4:3009pMiniT BsrGI+NheI 371&nbsp; &nbsp;NEB 2<br />
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Time:30min<br />
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'''Result:1727 can see band as vector, 945pminiT2 can see band as insert, cut out and combine,low concentration of 309pMiniT'''<br />
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[[File:Subclone of 945.JPG|RTENOTITLE]<br />
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Date:3/15/2018<br />
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'''3009pMiniT2 digestion test <span style="font-size:larger;">1</span>'''<br />
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Restriction enzymes:BsrG I+NheI<br />
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Buffer:NEB 2<br />
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Time:30min<br />
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'''Result:seems Lane 4 has band'''<br />
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[[File:IMG 3602.JPG|RTENOTITLE]]<br />
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'''3009pMIniT2 digestion test <span style="font-size:larger;">2</span>'''<br />
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Restriction enzymes:XhoI<br />
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Buffer:Fastdigest<br />
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Time:10min<br />
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7:3009-4pminiT2 (sequenced)<br />
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'''Result:some of them have bands, Lane 4 has band too.'''<br />
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[[File:3009pMIniT2 digestion test 2.JPG|RTENOTITLE]]<br />
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'''3009pMiniT2 digestion Test&nbsp;'''<span style="font-size:larger;">'''3'''</span><br />
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1 3009-4 BsrGI +KpnI<br />
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2 3009#5 BsrGI +KpnI<br />
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3 3009#5 BsrGI +NdeI<br />
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4 3009#5 NdeI +NheI<br />
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Buffer:NEB 2<br />
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Time:30min<br />
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[[File:3009pMiniT2 digestion Test 3.JPG|RTENOTITLE]]<br />
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== Analysis of&nbsp;3009-4 in pMiniT2 ==<br />
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[[Category:Cloning]]3009-4 in pMiniT2 digested with Nhe1-BsrG1, Kpn1-BsrG1<br />
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Date: 3/26/18, by Paul<br />
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Buffer: FastDigest<br />
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37 oC 1.5 hour<br />
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[[File:3009 4 BsrG1 Kpn1 Nhe1.JPG]]<br />
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don't know if Nhe1 sitte is present, need to transform INV110</div>
Crm