length limits of mutagenic primers
mb at laplace.csb.yale.edu
Wed Aug 9 08:48:51 EST 2000
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.
More information about the Methods