MORPH mutagenesis kit

A.S. Sohal u9310871 at MUSS.CIS.MCMASTER.CA
Wed Jun 19 12:30:08 EST 1996


Hi! This is the guy who was having trouble with the MORPH mutagenesis 
kit.  Well I've been able to isolate my plasmid from the mut S cells for 
some unknown reason but the problem now has become one of efficiency.  
Out of 24 tested clones, only 5 showed the creation of the mutation 
(which was the creation of a unique restriction endonuclease site).  Of 
those 5, only 1 gave the expected results while the other 4 gave some 
strange band sizes upon digestion and electrophoresis.  My supervisor has 
deemed that this type of efficiency (1 out of 24) is unacceptable since 
if used to make mutants that don't necessarily create a new restriction 
endonuclease site, screening will be a problem.  I was wondering if 
someone had any ideas why these results are so far off from the 
theoretical efficiency of 50%.  Any clues would be greatly appreciated.

Thanx

Avtar Sohal



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