I need to design primer for inverse PCR site directed mutagenesis where primers are not complementary to each other and anneal back to back at 5' end (not a single base is complementary in primer)
In attached Image I am interested for strategy (A) substitution of base by mutagenesis. So how to design primer for this strategy also provide detailed protocol for this kind of mutagenesis ?
What is the success rate for 1 bp substitution by using this mutagenesis strategy?
I am not getting whole information for this so please help .