Did you know that this isothermal amplification method can be performed in as little as 5-10 minutes with limited resources? Get a quick overview on how LAMP works in this animation.
When designing a LAMP experiment, four to six primers are required, specific to identified regions of target DNA or RNA sequence. Amplification initiates from strand invasion by one of the inner primers and a stand displacing DNA polymerase extends the primer and separates the target DNA duplex. The first product is then displaced by synthesis initiating from an outer primer, which anneals to an upstream target region. As it is displaced, the end of the product forms a self-hybridizing loop structure, due to inclusion of a reverse complementary sequence in the inner primer sequence. This annealing and displacement cycle repeats on the opposite end of the target sequence and the resulting product is a short dumbbell structure that forms a seed for exponential lamp amplification. This LAMP dumbbell structure contains multiple sites for initiation of synthesis from the three prime ends of the open loops and annealing sites for both the inner and loop primers. As amplification precedes from these multiple sites, the products grow and form long concatemers, each with more sites for initiation. The result is a rapid accumulation of double-stranded DNA and the amplification by-products that can be detected by a variety of methods.