The Mutagenesis reactions were performed by using 1U of TagMaster® Enzymes and 5ng of 7.4kb template plasmid under conditions of 95°C 5min, 20 cycles of (95°C 20sec, 60°C 10sec, 70°C 3min 20sec). The reaction products were run in a 1% agarose gel (as shown above). The products were then directly transformed into TagMaster® competent cells. The insertion of tags was verified by sequencing. The successful rates of all 6 mutagenesis reactions were 82%~91%.