Table 2.
Cq values obtained with ZNA, DNA and LNA primers under various MgCl2 concentrations and annealing temperatures
| Primer type | MgCl2 (mM) |
Cq target DNA |
Cq control DNA |
||||
|---|---|---|---|---|---|---|---|
| 57°C | 60°C | 63°C | 57°C | 60°C | 63°C | ||
| ZNA | 1.5 | 16.5 ∓ 0.10 | 16.4 ∓ 0.03 | 16.2 ∓ 0.06 | 27.4 ∓ 0.57 | 28.1 ∓ 0.04 | 34.4 ∓ 0.48 |
| 3 | 16.7 ∓ 0.21 | 16.9 ∓ 0.08 | n.d | 19.6 ∓ 0.03 | 22.3 ∓ 0.07 | n.d | |
| 4 | 16.5 ∓ 0.05 | n.d | n.d | 17.6 ∓ 2.10 | n.d | n.d | |
| DNA | 1.5 | 27.5 ∓ 0.10 | 30.7 ∓ 0.52 | 33.2 ∓ 0.32 | >45 | >45 | >45 |
| 3 | 20.4 ∓ 0.03 | 20.2 ∓ 0.11 | 21.8 ∓ 0.11 | 34.5 ∓ 1.09 | 35.4 ∓ 1.7 | 38.5 ∓ 1.23 | |
| 4 | 19.6 ∓ 0.00 | n.d | n.d | 31.0 ∓ 0.18 | n.d | n.d | |
| LNA | 1.5 | 24.6 ∓ 0.38 | 24.1 ∓ 0.04 | 24.0 ∓ 0.23 | 38.0 ∓ 1.41 | >45 | >45 |
| 3 | 20.0 ∓ 0.13 | 19.4 ∓ 0.12 | 19.1 ∓ 0.06 | 29.6 ∓ 0.6 | 32.6 ∓ 2.79 | 35.2 ∓ 1.51 | |
| 4 | 19.4 ∓ 0.06 | n.d | n.d | 28.5 ∓ 0.04 | n.d | n.d | |
Target genomic DNA (10 ng) or control DNA (10 ng) were amplified in duplicate with 100 nM of each E7 primer type as specified. All reactions were performed in duplicate (replicate standard deviation provided by the Rotor-Gene 6000 analysis software is indicated) using Sensimix NoRef kit 0.5×. MgCl2 was adjusted to the indicated concentration. Cycling conditions were 94°C for 20 s, annealing for 20 s, 72°C for 15 s.
n.d., not determined.