Morph Site Directed Mutag. Protocol

Stephen Bell spbell at mit.edu
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
ssDNA.







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