Problem with Site-Directed Mutagenesis

Todd DeZwaan DEZWAAN_T at a1.mscf.upenn.edu
Sat Apr 29 13:13:25 EST 1995


While creating a series of site-directed mutations in a specific region of
my gene of interest I have generated a mutant which gives two different
phenotypes; lethal and temperature sensitive.  The discrepancy can only be
linked to differences in the input, mutant construct and not to variations
in the host background for a number of reasons which I won't go into.  My
question is this, is it possible that I have a sub-population of mutagenic
oligonucleotides which would account for the presence of two different
mutant constructs in a single prep (i.e., what is the possibility that a
base was misincorporated in some of my mutagenic oligonucleotides during
synthesis) ?  I am using the Clonetech Transformer Site-Directed
Mutagenesis Kit for these experiments.  Thanks in advance for any help in
this matter.

Todd DeZwaan



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