cloning problems

Frosso Voulgaropoulou frossov at worldnet.att.net
Sat Aug 9 03:12:50 EST 1997


I have a cloning problem.  I had cloned PCR fragments into a tet-R 
vector using primers that I had engineered the BglII and PpuMI sites. 
PpuMI is a methylation sensitive site so I need to pass the DNA 
through methylation deficient bacterial strains (such as DM1 from 
BRL).  The problem I have is that I cannot transform DM1 cells 
(methylation deficient strain) with this plasmid whereas, the same 
plasmid can transform DH5a and HB101 cells very well.  In addition, 
DM1 cells can be transformed very efficiently with other plasmids.  
Has anyone heard about this before?


Frosso  Voulgaropoulou
frossov at worldnet.att.net



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