length limits of mutagenic primers

[Mark Bowen]

I generally put 12-15 bases on either side of the mutegenized region.
The mismatches are centered.  I never worry about getting the melting
temp higher than what arises from the above methodology.  I have made
dozens of mutants this way..  The largest region I've altered with this
method is 16bp (4 contiguous codons).  It would not likely work for much
larger than this.

--
Mark Bowen