Typical LAMP Protocol (NEB #M0402)

Incubate the following reaction at 65°C for 30–60 minutes.



Final Concentration

10X Isothermal Amplification Buffer (Lyo-compatible)

2.5 µl


dNTP Mix (10 mM)

3.5 µl

1.4 mM each

FIP/BIP Primers (25X)

1 µl

1.6 µM

F3/B3 Primers (25X)

1 µl

0.2 µM

LoopF/B Primers (25X)

1 µl

0.4 µM

Bst 2.0 WarmStart DNA Polymerase (Glycerol-free) (8,000 U/ml)

1 µl

320 U/ml

DNA Sample


> 10 copies

Nuclease-free Water

to 25 µl


Total Reaction Volume

 25 µl


 General Guidelines:

  1. A LAMP Primer Mix can be prepared with all 4 or 6 (with Loop) primers. A 25X Primer Mix should contain: 40 µM FIP, 40 µM BIP, 5 µM F3, 5 µM B3, 10 µM LoopF, 10 µM LoopB in TE or water.

  2. Dilute the Bst 2.0 WarmStart DNA Polymerase (Glycerol-free) (NEB #M0402) to 8,000 U/ml in 1X Isothermal Amplification Buffer (Lyo-compatible).

  3. If analyzing via agarose gel electrophoresis or other method requiring opening LAMP reaction vessels, set up a secondary analysis area and equipment to avoid contamination.

  4. Running a no-template control is strongly recommended to ensure amplification specificity.

  5. If optimization is desired, try titrating Bst 2.0 WarmStart DNA Polymerase (Glycerol-free) (0.04-0.32 U/µl), or changing reaction temperature (50–68°C).