Frequently Asked Questions

1. Should I store the TrueLAMP Kit at –20 °C? 
Yes. While the 2× buffer may be stored at 4–22 °C with the tube tightly capped, CO₂ can dissolve into the buffer over time and lower its pH, causing gradual color fading. Freezing slows this process.

2. How does TrueLAMP eliminate primer amplification in NTCs? 
The buffer contains a proprietary inhibitor formulated with the polymerase to prevent nonspecific primer amplification when no template is present.

3. What is the best incubator for TrueLAMP reactions? 
The reaction must be heated evenly to prevent evaporation in small volumes. A digital oven or submerged water bath or thermocycler with heated lid works best. 

4. Why am I still getting primer amplification in the NTC?
•    Warm the buffer before use. Cold storage (–20 to 4 °C) can form microcrystals that reduce inhibition.
•    Recalibrate pipettes. Incorrect volumes (< 4.75 µL buffer per 10 µL reaction) may also reduce inhibition.
•    Verify incubation temperature. Nonspecific amplification can occur at ≤ 63 °C. Temperature distribution in digital ovens may be uneven—calibrate and adjust placement of 8-strips as needed. For multiple strips, use a water bath.
•    Strengthen inhibition. Increase buffer slightly (e.g., 5.25 µL 2× buffer per 10 µL reaction) for particularly stubborn primer sets.

5. Why was my positive control not amplified?
•    Determine the limit of detection (LOD) for the primer set. Template concentration < LOD will not be amplified consistently.

6. Can I use raw samples (e.g., saliva, plasma)? 
Not recommended. TrueLAMP is sensitive to pH and ionic strength. For best results, use purified DNA or RNA. Although water-suspended virions may be used directly with TrueLAMP, its compatibility with other types of cells has not been fully characterized. 

7. Can I pre-mix polymerase into the 2× buffer?
No. The polymerase is inactivated within hours, even at –20 °C. Always prepare the reaction mix fresh.

8. How should I quantitate the color change following LAMP amplification?
Color intensity of the reaction may be quantified using a smartphone colorimeter app with its smallest aperture by sampling the region at the center immediately below the liquid meniscus and recording the maximum magenta (CMYK mode) or minimum green (RGB mode).