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.