Fig. 5

Effect of cycling temperatures on the efficiency of gene synthesis. Panel a: different annealing temperatures were studied (1: 50 °C; 2: 52.3 °C; 3: 54.6 °C; 4: 59.6 °C and 5: 62 °C). Panel b: 22, 24 and 26 cycles combined with different times of denaturation, annealing and extension were used to synthesise gene C (I: 95 °C for 20 s, 60 °C for 10 s, 70 °C for 1 s; II: 95 °C for 16 s, 60 °C for 10 s, 70 °C for 1 s; III: 95 °C for 20 s, 60 °C for 8 s, 70 °C for 1 s; IV: 95 °C for 16 s, 60 °C for 8 s, 70 °C for 1 s; V: 95 °C for 20 s, 60 °C for 10 s, 70 °C for 3 s; VI: 95 °C for 16 s, 60 °C for 10 s, 70 °C for 3 s; VII: 95 °C for 20 s, 60 °C for 8 s, 70 °C for 3 s; VIII: 95 °C for 16 s, 60 °C for 8 s, 70 °C for 3 s). The extension times used for each number of cycles were 1 and 3 s