Popular lifehacks

What is the principle of loop-mediated isothermal amplification?


What is the principle of loop-mediated isothermal amplification?

Loop-mediated Isothermal amplification (LAMP), is an emerging technology that allows DNA amplification at a constant temperature. The key to this principle is the use of a DNA polymerase that possesses strand displacement activity.

How does loop-mediated amplification work?

Loop-mediated isothermal amplification employs a DNA polymerase with strand-displacement activity, along with two inner primers (forward and backward inner primers) and two outer primers (F3 and B3) that recognize six separate regions within a target DNA sequence.

How does LAMP assay work?

In a LAMP assay, the reaction takes place in a single tube containing buffer, target DNA, DNA polymerase and primers. The amplified product can be detected by naked eye as a white precipitate or a yellow-green color solution after addition of SYBR green to the reaction tube.

What is the principle of LAMP?

Loop-mediated isothermal amplification (LAMP), a newly developed gene amplification method, combines rapidity, simplicity, and high specificity. Several tests have been developed based on this method, and simplicity is maintained throughout all steps, from extraction of nucleic acids to detection of amplification.

What is a loop primer?

The Loop Primers (either Loop Primer B or Loop Primer F), containing sequences complementary to the single stranded loop region (either between the B1 and B2 regions, or between the F1 and F2 regions) on the 5′ end of the dumbbell-like structure, provide an increased number of starting points for DNA synthesis for the …

What is PCR LAMP for?

Loop-mediated isothermal amplification (LAMP) is a single-tube technique for the amplification of DNA and a low-cost alternative to detect certain diseases.

Is LAMP a type of PCR?

However, LAMP differs from PCR in several ways. It uses four to six primers to recognize six distinct regions of DNA or RNA, while PCR uses two primers to recognize two regions. The primers in LAMP cause DNA strand displacement and causes the end of the DNA strand to form a loop.

What is PCR What does it do?

PCR means polymerase chain reaction. It’s a test to detect genetic material from a specific organism, such as a virus. The test detects the presence of a virus if you have the virus at the time of the test. The test could also detect fragments of the virus even after you are no longer infected.

Why do we need loop mediated isothermal amplification ( LAMP )?

Moreover, LAMP excels through its isothermal and energy efficient amplification requirements, rendering it a prime candidate for low-cost diagnostics and analysis at the point of need.

How is isothermal amplification used in real time detection?

The complex and expensive devices required for thermal cycling and real-time detection during PCR restrict the use of this amplification method. Isothermal NAATs enable amplification reactions at constant and moderate temperatures.

How many primers are needed for loop mediated amplification?

Loop-mediated isothermal amplification (LAMP) is well known for its robust and highly sensitive and specific amplification of target DNA, which is achieved by utilizing up to six primers.

Why are isothermal nucleic acid amplification tests important?

In the course of the last 20 years, isothermal nucleic acid amplification tests have emerged as an important diagnostic tool, not only for clinical applications, but also for food quality control and environmental monitoring.