Table 2 The amplification conditions used in Touchdown PCR for the construction of 3 fragments of the human IGF-IR gene promoter (∞: hold)

1

Denature

95°C

15 min

2

Denature

95°C

30 sec

3

Anneal

70°C

45 sec

−1.0°C^*

4

Elongate

72°C

1 min

Phase 2

Step

Temperature

Time

5

Denature

95°C

30 sec

6

Anneal

60°C

45 sec

7

Elongate

72°C

1 min

Termination

Step

Temperature

Time

8

Elongate

72°C

5 min

9

Halt reaction

4°C

15 min

10

Hold

4°C

∞