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3131 pCEL0-HA-VL-mIgKappa - Revision history
2026-10-18T14:45:04Z
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Crm at 17:19, 21 March 2018
2018-03-21T17:19:19Z
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<p><b>New page</b></p><div>Subclone of 3131:<br />
<br />
Restriction enzymnes:BssHII+XhoI<br />
<br />
Time:30min<br />
<br />
Buffer:Fastdigest<br />
<br />
result:good, combine and do ligate<br />
<br />
[[File:Subclone of new 3131.JPG|RTENOTITLE]]<br />
<br />
'''Miniprep digestion of 3131_3161_3167'''<br />
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Name length Enzymen Buffer<br />
<br />
3131:1-4 342 BssHII+XhoI Cutsmart<br />
<br />
3161:5-8 225 NdeI+XhoI Cutsmart<br />
<br />
3167:9-12 397 BsrGI+NheI NEB 2<br />
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Time:40min<br />
<br />
[[File:Miniprepof 3131 3167 3161.JPG|RTENOTITLE]]<br />
<br />
'''Redo miniprep of 3131_3161_3167'''<br />
<br />
3131:1-4 BssHII+XhoI 342 Cutsmart<br />
<br />
3161:5-8 NdeI+XhoI 225 Cutsmart<br />
<br />
3167:9-12 BsrGI+NheI 397 NEB 2<br />
<br />
Time:40min<br />
<br />
[[File:Redo miniprep of 3131 3161 3167.JPG|RTENOTITLE]]<br />
<br />
Date:3/15/2018<br />
<br />
'''MiniPrep digestion of 3131'''<br />
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Enzymes:BssHII+XhoI<br />
<br />
Buffer:Cutsmart<br />
<br />
Time:40min<br />
<br />
'''Result:2,3,4,seems good, do sequencing,select #3'''<br />
<br />
[[File:Miniprepdigstion3131.JPG|RTENOTITLE]]<br />
<br />
'''Miniprep of 947_947pMiniT2_3131_3152'''<br />
<br />
947:1-4 XbaI+NcoI-HF 1394 Cutsmart<br />
<br />
947pMiniT2:5-8 ECoRI Cutsmart<br />
<br />
3131:9-12 BssHII+XhoI 342 Cutsmart<br />
<br />
3152:13-16 BsrGI+KpnI 371 NEB 2<br />
<br />
'''Result: sent lane 2, lane 16 to do sequence'''<br />
<br />
[[File:IMG 3621.JPG|RTENOTITLE]]<br />
<br />
Date:3/19/2018<br />
<br />
'''Redo Subclone'''<br />
<br />
'''3131''':BssHII+XhoI<br />
<br />
Buffer:Cutsmart<br />
<br />
'''Results:cut out the middle band of insert, and cut out the vector.'''<br />
<br />
[[File:IMG 3626.JPG|RTENOTITLE]]<br />
<br />
'''Date:3/21/2018'''<br />
<br />
'''Miniprep of 3165pMiniT2_3165_3131'''<br />
<br />
3165pMiniT2:1-5 EcoRI<br />
<br />
3165:6-10 BsrGI+NdeI<br />
<br />
3131:11-15 BssHII+XhoI<br />
<br />
Time:30min<br />
<br />
'''Result:3165pMiniT2(1-5) have bands,3131(12-15)have bands.redo 3165 digestion.'''<br />
<br />
<br />
<br />
[[File:IMG 3637.JPG]]<br />
[[Category:Cloning]]</div>
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