Fig 2. Successful insertion of different tags (6His, HA, Myc+Flag, Myc, Flag) into a 7.4kb plasmid.

The Mutagenesis reactions were performed by using 1U of TagMaster Enzymes and 5ng of 7.4kb template plasmid under conditions of 95°C 5min, 20cycles of (95°C 20sec, 60°C10sec, 70°C 3min20sec). The reaction products were run in 1% agrose 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%.