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Related Experiment Videos

Loop-mediated isothermal amplification of DNA.

T Notomi1, H Okayama, H Masubuchi

  • 1Eiken Chemical Co. Ltd, 1381-3 Shimoishigami, Ohtawara, Tochigi 324-0036, Japan, Department of Biochemistry and Molecular Biology, The University of Tokyo, Graduate School of Medicine, Bunkyo-ku, Tokyo 113-0033, Japan. tsugunori_notomi@eiken.co.jp

Nucleic Acids Research
|June 28, 2000
PubMed
Summary

We developed loop-mediated isothermal amplification (LAMP), a novel DNA amplification method. LAMP achieves high specificity, efficiency, and speed under isothermal conditions, making it ideal for rapid DNA detection.

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Area of Science:

  • Molecular Biology
  • Biotechnology

Background:

  • DNA amplification is crucial for molecular diagnostics and research.
  • Existing methods like PCR require thermal cycling, limiting their field applicability.
  • There is a need for rapid, specific, and efficient DNA amplification under isothermal conditions.

Purpose of the Study:

  • To develop a novel DNA amplification method.
  • To achieve high specificity, efficiency, and rapidity under isothermal conditions.
  • To enable rapid DNA detection without thermal cycling.

Main Methods:

  • Developed loop-mediated isothermal amplification (LAMP).
  • Utilized a DNA polymerase and four specific primers recognizing six target DNA sequences.
  • Initiated amplification with an inner primer, followed by strand displacement synthesis.

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  • Employed cycling with inner primers hybridizing to stem-loop structures for exponential amplification.

    • Main Results:

    • Achieved amplification of 10(9) target copies in under an hour.
    • Generated stem-loop DNA products with multiple inverted repeats.
    • Demonstrated high selectivity due to recognition of multiple distinct sequences.
    • Amplification occurred under isothermal conditions, eliminating the need for thermal cycling.

    Conclusions:

    • Loop-mediated isothermal amplification (LAMP) is a highly specific, efficient, and rapid DNA amplification technique.
    • LAMP operates under isothermal conditions, offering advantages for point-of-care and field applications.
    • The method's reliance on multiple primer recognition sites ensures high target selectivity.