Universal Multiplex PCR: a novel method of simultaneous amplification of multiple DNA fragments

Table 2 The UM-PCR procedure (“Two Rounds Mode”)

	Temperature (°C)	Time
	94	5 min
	94	40s
	annealing temperature of primer 1	20s
	annealing temperature of primer 2	20s
	annealing temperature of primer 3	20s
	annealing temperature of primer 4	20s
	annealing temperature of primer 5	20s
	72	30s
	94	40s
	70	50s
	72	10 min

In the first round (the first three cycles), annealing temperatures that are optimized in accordance with the Tm of normal SSR primers are sorted in descending order. Annealing temperatures may be different in various combinations of U-primers.

ISSN: 1746-4811