Morph Site Directed Mutag. Protocol

Stephen Bell spbell at
Fri Jan 26 09:52:47 EST 1996

 Has anyone tried the Morph Protocol from 5'->3' for site directed
mutagenesis or a protocol like it?  To avoid the usual ssDNA production
in a dut- ung- strain, they melt double stranded methylated DNA and
anneal a mutagenic primer.  The primer is extended by T4 DNA polymerase
and the reulting DNA is cut with Dpn I (only cuts methylated and not
hemi-methylated DNA) and transform into a mutS strain allowing
production of both the wild type and mutant strands of DNA. Seems simple
enough and all one really needs is the mutS strain, no helper phage, no

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