3021 pCEL0-mIg-LCKappa

PCR from  IgG kappa LC from 390	SI1-Unc5b

3021-Xba1-BssHII-F



     Xba1     MssHII             KpnI        Xho1     
atcatc TCTAGA  GCGCGC atc GGCTCC  GGtACC AAG CTCGAG ATC AAa CGC GCTGATGCTG
                           G  S    G  T   K   L  E   I   K   R   A  D  A 

3021-Age1-Xba1-R
atcatcTCTAGAACCGGTGCGGCCGCCTAACACTCATTCCTGTTGAAG
                          Not1     Age1    Xba1
  (CTTCAACAGGAATGAGTGTTAG GCGGCCGC ACCGGT TCTAGA gatgat )

Vector:   MssHII  - Nhe1
insert:  MssHII  - Xba1

in pMiniT2 vector digestion with Xho1

date:1/31/2018 by Yijia

374 bp

Miniprep digestion:

3021pMiniT2:1-4

Restriction enzymes:EcoRI

3021:1-4 restriction enzymes:BssHII+XbaI

3027:5-8 restrcition enzymes:NotI+NheI

Buffer:Fastdigest

Time:16min

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.

miniprep digestion 3021

3021pMiniT2:BssHII+XbaI 374

3021:BssHII+XbaI 374 Cutsmart

3165:SalI+BsrGI 702 NEB 3

Time:30min