Method of synthesizing Nucleic Acid [LAMP]

In the Public Domain since

9 Nov 2018

Description

In 1998, Notomi Tsugunori et al., patented Loop-mediated Isothermal amplification (LAMP), an invention to provide a method capable of efficiently synthesising nucleic acid in a sequence specific manner at low cost and with high specificity using a single strand-displacing DNA polymerase enzyme e.g. Bst DNA polymerase, under isothermal reaction conditions at about 65°C.

This invention overcame some limitations of PCR (Polymerase Chain Reaction), the most popular technique of amplifying nucleic acid in vitro:

  • PCR requires special equipment such as a thermocycler to cycle temperatures in order to denature double stranded into a single stranded form, LAMP uses strand-displacing enzymes removing the need for this step.
  • PCR has a reaction time of 1-2 hours with LAMP can amplify DNA 109-1010 times in 15-60 minutes
  • LAMP reduces non-specific binding in the early stages of the reaction by including 4 primers to recognize 6 distinct regions of DNA.

The amplification product for LAMP is a looped structure consisting of alternately inverted repeats of the target sequence on the same strand.

For more information on LAMP see the Eiken Chemical Company website.

Abstract

The present invention relates to an oligonucleotide having a novel structure and a method of synthesizing nucleic acid by using the same as a primer. This oligonucleotide is provided at the 5'-side of the primer with a nucleotide sequence substantially the same as a region synthesized with this primer as the origin of synthesis. The present invention realizes the synthesis of nucleic acid-based on an isothermal reaction with a simple constitution of reagents. Further, the present invention provides a method of synthesizing highly specific nucleic acid on the basis of this method of synthesizing nucleic acid.

Category

Others > COVID-19

Related Inventors

unknown

Source

US Patent No 6,410,278

Patent Title: Method of synthesizing Nucleic Acid

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