LAMP (Loop-mediated isothermal amplification) is a technique for a quick amplification of nucleic acids. In contrast to the PCR technique, in which the reaction is carried out with a series of alternating temperature cycles, the LAMP reaction is carried out in a constant temperature of 65°C. The LAMP RT technique uses reverse transcriptase, which transcribes the RNA strand into cDNA, and Bst polymerase, which amplifies the resulting fragment. The reaction is exceptionally quick and efficient thanks to using a set of three pairs of primers specific for cDNA. In the first phase, a base product with loop formation at the ends is created, next the strand displacement activity and attachment of subsequent primers lead to the creation of increasingly longer DNA fragments consisting of the base product repeats. In the electrophoretic separation the obtained product is not a single band but constitutes a mixture of products of various sizes with repeated sequence. The detection is done by fluorescence, which allows for the measurement in real time. During the reaction, an insoluble precipitate of magnesium pyrophosphate is formed, which can be observed as turbidity.
Flo-LAMP RT Kit
cat. no. E1425