2X Fluorescent RT-LAMP Master Mix
Croyez 2X Fluorescent RT-LAMP Master Mix is an optimized master mix for reverse-transcription loop-mediated isothermal amplification (RT-LAMP) reactions. This product is a dual enzyme system, providing a rapid and sensitive detection in one pot. A fluorescent dye is also supplied with the kit. The LAMP reactions can be monitored through real-time fluorescence detection.
The following procedure is a general guideline for RT-LAMP reaction. To maintain an RNase-free environment, always wear disposable gloves, and use laboratory consumables and water of nuclease-free grade during the whole experiment course.
RT-LAMP reaction set-up:
1.10X LAMP primer mix
2. An overview of the reaction set-up is listed in the table below. Place all required reagents on ice. Distribute appropriate volumes into each tube before adding template.
* See Usage Notes for additional guidelines on primer/template preparation.
3. Add target RNA template to the detection tube. Gently mix the reaction thoroughly to achieve uniform distribution and avoid making bubbles.
4. Incubate at 65°C for 30-60 min.
5. After LAMP reaction complete, the enzyme can be inactivated by heating at 80°C for 10 min.
6. For real-time detection, collect fluorescent data using the SYBR® or FAM channels.
Shipping Conditions:
Blue ice
RT-LAMP reaction set-up:
1.10X LAMP primer mix
| Component | 10X concentration | Final concentration |
| FIP | 16 μM | 1.6 μM |
| BIP | 16 μM | 1.6 μM |
| F3 | 2 μM | 0.2 μM |
| B3 | 2 μM | 0.2 μM |
| LOOP F | 8 μM | 0.8 μM |
| LOOP B | 8 μM | 0.8 μM |
2. An overview of the reaction set-up is listed in the table below. Place all required reagents on ice. Distribute appropriate volumes into each tube before adding template.
| Component | Amount | Final concentration |
| 2X Fluorescent RT-LAMP Master Mix | 12.5 μL | 1X |
| 10X LAMP primer mix | 2.5 μL | 1X |
| 50X LAMP Fluorescent Dye | 0.5 μL | 1X |
| RNA template | 1-2 μL | variable |
| Nuclease-Free H2O | X μL | - |
| Total reaction volume | 25 μL | - |
3. Add target RNA template to the detection tube. Gently mix the reaction thoroughly to achieve uniform distribution and avoid making bubbles.
4. Incubate at 65°C for 30-60 min.
5. After LAMP reaction complete, the enzyme can be inactivated by heating at 80°C for 10 min.
6. For real-time detection, collect fluorescent data using the SYBR® or FAM channels.
Shipping Conditions:
Blue ice
