TABLE 4.
Master mix components for serovar-specific monoplex PCR assays
Serovar | Concn (μM) of: |
PCR buffera | MgCl2 concn (mM)b | UNG amt (U) | |||
---|---|---|---|---|---|---|---|
Forward primer | Reverse primer | Probe 1 | Probe 2 | ||||
1 | 0.3 | 0.5 | 0.2 | 2 | 0.5 | ||
2 | 0.5 | 0.5 | 1 | 3.0 | 0.5 | ||
3 | 0.25 | 0.25 | 1 | 3.0 | 0.5 | ||
4 | 0.5 | 0.5 | 0.2 | 0.2 | 2 | 0.5 | |
5 | 0.4 | 0.5 | 0.2 | 2 | 0.5 | ||
6 | 0.4 | 0.4 | 1 | 3.0 | 0.5 | ||
7 | 0.25 | 0.25 | 1 | 3.0 | 0.5 | ||
8 | 0.25 | 0.25 | 1 | 3.0 | 0.5 | ||
9 | 0.25 | 0.25 | 1 | 3.0 | 0.5 | ||
10 | 0.3 | 0.5 | 0.15 | 2 | 0.5 | ||
11 | 0.2 | 0.5 | 0.2 | 2 | 0.5 | ||
12 | 0.5 | 0.5 | 0.2 | 0.2 | 2 | 0.5 | |
13 | 0.4 | 0.4 | 1 | 3.0 | 0.5 | ||
14 | 0.5 | 0.5 | 1 | 3.0 | 0.5 |
1, 2 μl LightCycler FastStart DNA Master SYBR green I; 2, 4 μl LightCycler FastStart DNA MasterPLUS HybProbe.
Final MgCl2 concentration.