WarmStart Colorimetric LAMP 2X Master Mix Typical LAMP Protocol (M1800)

Reaction Setup: For simplicity in setting up reactions, we recommend making stocks of the LAMP primers at a usable concentration. For example, we suggest a 10X Primer Mix containing all 6 LAMP primers.
A 10X LAMP Primer Mix contains:
 FIP  16 μM
 1.6 μM
 BIP  16 μM  1.6 μM
 F3  2 μM  0.2 μM
 B3  2 μM  0.2 μM
 4 μM  0.4 μM
 4 μM  0.4 μM

Note: Make primer stock in molecular biology grade H2O rather than TE or other buffer in order to avoid carryover of additional buffer to the LAMP reaction. Prepare primer stocks in nuclease free water and store at –20°C for up to 2 years.

1.    Thaw all components to be used at room temperature and place on ice. Salt may appear in the bottom of the tube so vortex briefly or invert tubes several times to mix thoroughly. Centrifuge to collect material and place on ice.

2.    Prepare reaction mix as described below using Colorimetric LAMP Master Mix, LAMP primers and nuclease free water. Volumes listed per 25 μl LAMP reaction, but other volumes (10, 20, 50 μl etc.) are all effective if desired, just adjust volumes accordingly. Sample is assumed here to be 1 μl, but for higher sample volumes add as needed and reduce volume of H2O to compensate. For non-template reactions add equivalent volume of H2O or sample storage buffer.


WarmStart Colorimetric LAMP 2X Master Mix
 12.5 µl
 12.5 µl
 12.5 µl
LAMP Primer Mix (10X)
 2.5 µl
 2.5 µl
 2.5 µl
Target DNA
 1 µl
 –  –
Target RNA
 –  1 µl
dH2O  9 µl
 9 µl
 10 µl
Total Volume
 25 µl
 25 µl
 25 µl

3.    Vortex reaction mix and centrifuge to collect material.

4.    Pipet 24 μl per reaction into desired reaction vessels and add 1 µl of sample. Mix by vortexing or by pipetting if using a plate or similar vessel, centrifuge to collect if necessary. Check that reaction solutions have a bright pink color, which indicates initial high pH required for successful pH-LAMP reaction.

5.    Seal reaction vessels.

6.    Incubate at 65°C for 30 minutes.  

7.    Remove tubes or vessels from incubation and examine by eye. Positive reactions will have turned yellow while negative controls should remain pink. If color change is not robust, e.g. an orange color is visible, return reactions to 65°C for an additional 10 minutes. Reactions can be examined earlier if desired, and high copy or input reactions can exhibit full color change in as little as 10–15 minutes. Color will be visible directly on removal from incubation temperature, but can be intensified by allowing reaction to cool to room temperature.

8.    The result can be photographed or scanned to record the colorimetric results, or simply kept at room temperature in the reaction vessel.