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    Nucleic acid amplification is an important process in biological systems and is the leading method used to detect and analyze small amounts of nucleic acids. Polymerase chain reaction (PCR) is currently the most widely used method for DNA amplification. However, it requires a thermal cycling machine to separate two DNA strands and then amplify the desired fragment. Isothermal amplification of nucleic acids is a simple process that allows rapid and efficient accumulation of nucleic acid sequences at a constant temperature. As an alternative to PCR, various isothermal amplification techniques have been developed. Isothermal amplification may sometimes have lower sensitivity and/or specificity than PCR but is ideal for situations where thermal cyclers are not available, and is faster, simpler, and more cost-effective. Based on different enzymatic mechanisms, isothermal amplification can be divided into four main categories.

    • Nucleic acid sequence-based amplification (NASBA) and signal-mediated amplification of RNA technology (SMART) are two isothermal amplification technologies based on elements of RNA transcription. NASBA amplifies the target nucleic acid directly from RNA or DNA via complementary DNA (cDNA) intermediates. The SMART assay that relies on signal amplification generates an RNA signal from the target nucleic acid and subsequently uses this RNA signal for the detection or to generate additional RNA signals.
    • Helicase-dependent amplification (HDA) and recombinase polymerase amplification (RPA) are isothermal amplification technologies based on DNA replication with enzymatic double-stranded fusion/in situ annealing.
    • Isothermal amplification technologies based on DNA-polymerase-mediated strand displacement from linear/circular targets include rolling circle amplification (RCA) and loop-mediated isothermal amplification (LAMP). These methods use DNA polymerases with strand replacement capabilities to produce continuously growing nucleic acids in a self-replacing loop.
    • Other isothermal amplification techniques are based on polymerase extension/strand replacement and single-strand cleavage events, such as strand displacement amplification (SDA). These techniques are performed using a restriction endonuclease incision followed by polymerase replacement of the incised strand as a new strand is extended from the incision site.

    Amerigo Scientific offers a wide range of isothermal amplification reagents and kits that can be used in bioanalytical, nanotechnology, materials science, and device integration applications.

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    • Catalog Number: IAR2465224DNA
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    • Catalog Number: IAR2388544QIT
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