An example of making multiple mutations across a wide region. (A) A fragment of the human nAChR α2 subunit cDNA with 8 arginine (R, in bold) codons (top) to be substituted by glutamine (Q) codons (bottom). Mutated nucleotides are indicated in red. (B) Actual forward (45-base) and reverse (46-base) primers with 16 bp overlapping in their 5' ends.
Li et al. BMC Biotechnology 2011 11:92 doi:10.1186/1472-6750-11-92