Table IV.

TAIL-PCR cycling parameters used to isolate flanking DNA from insertional mutants

Reaction	Step	Thermal Settings	No. of Cycles
Primary	1	95°C, 2 min	1
	2	94°C, 1 min; 62°C, 1 min; 72°C, 2.5 min	5
	3	94°C, 1 min; 25°C, 3 min; ramping to 72°C over 3 min; 72°C, 2.5 min	1
	4	94°C, 30 s; 68°C, 1 min; 72°C, 2.5 min; 94°C, 30 s; 68°C, 1 min; 72°C, 2.5 min; 94°C, 30 s; 44°C, 1 min; 72°C, 2.5 min	15
	5	72°C, 5 min	1
Secondary	1	94°C, 30 s; 64°C, 1 min; 72°C, 2.5 min; 94°C, 30 s; 64°C, 1 min; 72°C, 2.5 min; 94°C, 30 s; 44°C, 1 min; 72°C, 2.5 min	12
	2	72°C, 5 min	1
Tertiary	1	94°C, 30 s; 44°C, 1 min; 72°C, 2.5 min	20
	2	72°C, 5 min	1