Table 2 Primer Design Parameters Used in the Four Passes of Primer3.
First Pass | Second Pass | Third Pass | Fourth Pass | |
|---|---|---|---|---|
Optimum Size (BP) | 25 | 25 | 25 | 25 |
Minimum Size (BP) | 20 | 20 | 18 | 20 |
Maximum Size (BP) | 30 | 30 | 33 | 30 |
OptimumTM (°C) | 60 | 60 | 60 | 60 |
Minimum TM (°C) | 55 | 55 | 50 | 55 |
Maximum TM (°C) | 65 | 65 | 70 | 65 |
Optimum GC% | 50 | 50 | 50 | 50 |
Minimum GC% | 40 | 40 | 35 | 40 |
Maximum GC% | 60 | 60 | 65 | 60 |
GC CLAMP | Yes | No | No | Yes |
NUM RETURN | 1 | 1 | 1 | 1 |
PRODUCT SIZE RANGE (BP) | 400-600 | 400-600 | 400-600 | 400-600 |
Repeat Masking | Yes | Yes | Yes | No |
SNP Masking | Yes | Yes | Yes | Yes |