Table 5. Real-time and colorimetric reaction mixes of the mismatch-tolerant RT-LAMP assay.
| Component | Real-time RT-LAMP (μl) | Colorimetric RT-LAMP (μl) |
| 10× Isothermal Amplification Buffer | 2.5 | - |
| WarmStart Colorimetric LAMP 2x Master Mix | - | 12.5 |
| MgSO4 (100 mM) | 1 | - |
| dNTPs (10 mM) | 3.5 | - |
| Bst 2.0 DNA polymerase (8 U) | 1 | - |
| Warmstart RTx reverse transcriptase (15 U) | 0.5 | - |
| Q5 high-fidelity DNA polymerase (0.2 U/μl) | 0.75 | 0.75 |
| SYTO 9 (1×) | 1 | - |
| Mixed primer set 1 (25×) | 1 | 1 |
| Mixed primer set 2 (25×) | 1 | 1 |
| Mixed primer set 3 (25×) | 1 | 1 |
| RNA template | 3 | 3 |
| Nuclease-free water | 8.75 | 5.75 |
| Total | 25 | 25 |
Note: Obtain 0.2 U/μl Q5 high-fidelity DNA polymerase by ten-fold dilution of 2 U/μl Q5 enzyme stock.