Selection Problem in pUC18

Yeon Chul Kim yckim at chiak.kaist.ac.kr
Mon Oct 16 16:28:48 EST 1995


Hello!

pUC18 molecules were cut with SmaI and treated with CIP.  Taget DNA was

PCR product.  After ligation with T4 DNA ligase, I performed 

electroporation into XL1-Blue.  After transformation step, those cells

were plated on LB/Amp/Tet/IPTG/X-gal plate.  And I got some white

colonies.  I sequenced some DNAs.  And I got a strange result.

That sequence was same as pUC18 except deleting CCGG at SmaI site.
               
[---AGG ATC C(CC GG)G TAC CGA---]
              ^^ ^^
I'm wonder why I got that result.

Does anyone have answer?  Please let know.

Yeon Chul Kim

Bioprocess Engineering Research Center
KAIST, Korea
 




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