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  • Nested PCR in magnetically actuated circular closed-loop PCR microchip system

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    Author(s)
    Lok, Khoi Seng
    Lee, Peter Peng Foo
    Kwok, Yien Chian
    Nam-Trung, Nguyen
    Griffith University Author(s)
    Nguyen, Nam-Trung
    Year published
    2012
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    Abstract
    We demonstrate a new and sensitive amplification technique (referred to as Nested Polymerase Chain Reaction; nPCR). It based on a magnetically actuated circular closed-loop PCR microchip system. nPCR involves the use of two sets of primers in two successive PCR runs, and allows the amplification of a single locus from a minute quantity of template DNA. Two sets of primers are specially designed to a target 500-bp region of the bacteriophage lambda template DNA in the first PCR run, and a 247-bp region of the targeted 500-bp first PCR product in the second PCR run. PCR is run on the microchip system and concurrently in regular ...
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    We demonstrate a new and sensitive amplification technique (referred to as Nested Polymerase Chain Reaction; nPCR). It based on a magnetically actuated circular closed-loop PCR microchip system. nPCR involves the use of two sets of primers in two successive PCR runs, and allows the amplification of a single locus from a minute quantity of template DNA. Two sets of primers are specially designed to a target 500-bp region of the bacteriophage lambda template DNA in the first PCR run, and a 247-bp region of the targeted 500-bp first PCR product in the second PCR run. PCR is run on the microchip system and concurrently in regular thermocycler for comparison. The products are analyzed by conventional agarose gel electrophoresis. The detection limit for the initial template DNA is 1.63?׿105 copies per 匠(or 8.67 pg) for the first PCR run, and 1.63 copies per 匠(or 0.0867 fg) for the second run. The results are comparable to a regular thermocycler. This preliminary study opens a new gateway to future development of specialized nPCR on chip.
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    Journal Title
    Microchimica Acta
    Volume
    177
    Issue
    1-2
    DOI
    https://doi.org/10.1007/s00604-012-0760-2
    Copyright Statement
    © 2012 Springer Vienna. This is an electronic version of an article published in Microchimica Acta, Volume 177, Issue 1-2, pp 111-117, 2012. Microchimica Acta is available online at: http://link.springer.com// with the open URL of your article.
    Subject
    Analytical chemistry
    Instrumental methods (excl. immunological and bioassay methods)
    Other chemical sciences
    Microelectromechanical systems (MEMS)
    Publication URI
    http://hdl.handle.net/10072/50214
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    • Journal articles

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