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3021 pCEL0-mIg-LCKappa - Revision history
2026-10-18T05:51:24Z
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Crm at 15:16, 15 March 2018
2018-03-15T15:16:49Z
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<p><b>New page</b></p><div><pre>PCR from IgG kappa LC from 390 SI1-Unc5b<br />
<br />
3021-Xba1-BssHII-F<br />
<br />
<br />
<br />
Xba1 MssHII KpnI Xho1 <br />
atcatc TCTAGA GCGCGC atc GGCTCC GGtACC AAG CTCGAG ATC AAa CGC GCTGATGCTG<br />
G S G T K L E I K R A D A <br />
<br />
3021-Age1-Xba1-R<br />
atcatcTCTAGAACCGGTGCGGCCGCCTAACACTCATTCCTGTTGAAG<br />
Not1 Age1 Xba1<br />
(CTTCAACAGGAATGAGTGTTAG GCGGCCGC ACCGGT TCTAGA gatgat )<br />
<br />
Vector: MssHII - Nhe1<br />
insert: MssHII - Xba1</pre><br />
<br />
in pMiniT2 vector digestion with Xho1<br />
<br />
date:1/31/2018 by Yijia<br />
<br />
<span style="color: rgb(0, 0, 0); font-family: Arial, Verdana, sans-serif; font-size: 12px;">374 bp</span><br />
<br />
[[File:3130 3129 3021 3075.JPG|RTENOTITLE]]<br />
<br />
'''Miniprep digestion:'''<br />
<br />
3021pMiniT2:1-4<br />
<br />
Restriction enzymes:EcoRI<br />
<br />
3021:1-4 restriction enzymes:BssHII+XbaI<br />
<br />
3027:5-8 restrcition enzymes:NotI+NheI<br />
<br />
Buffer:Fastdigest<br />
<br />
Time:16min<br />
<br />
'''Result:'''seems good, but the result of sequencing is not,the product were actually are '''undigested vectors,3016pCEL0,3017pCEH0. so we collected them for other using.'''<br />
<br />
[[File:Miniprep Digestion 3021 3027.JPG|RTENOTITLE]]<br />
<br />
'''miniprep digestion 3021'''<br />
<br />
3021pMiniT2:BssHII+XbaI 374<br />
<br />
3021:BssHII+XbaI 374 Cutsmart<br />
<br />
3165:SalI+BsrGI 702 NEB 3<br />
<br />
Time:30min<br />
<br />
[[File:Redominiprep diegstion 3021.JPG|RTENOTITLE]]<br />
[[Category:Cloning]]</div>
Crm