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KMID : 0613820140240040428
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Journal of Life Science
2014 Volume.24 No. 4 p.428 ~ p.436
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Loop-mediated Isothermal Amplification (LAMP) for Detection of Streptococcus parauberis
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Moon Kyung-Mi
Kim Dong-Hwi
Heo Moon-Soo
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Abstract
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Loop-mediated isothermal amplification (LAMP) technique relies on autocycling strand displacement DNA sysnthesis without template denaturation steps under isothermal conditions. LAMP has more advantages than modern PCR, as it requires only basic laboratory equipment like an isothermal water bath, oven, and heating cabinet. Hence, in this study, five random primers were designed with Streptococcus parauberis, shikimate kinase Arok gene sequences (Genbank accession number: CP0024711). Two primers were selected based on the ladder pattern. Other optimum reaction conditions like temperature, time, and sensitivity were established and confirmed with conventional SYBR-green PCR.
Results confirmed that the designed random primers were species specific, without any non-target DNA amplification under isothermal conditions. Hence, this study suggests that LAMP techniques could be used in the diagnosis of fish pathogen, specifically S. parauberis.
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KEYWORD
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Loop-mediated isothermal amplification (LAMP), PCR, random primer, Streptococcus parauberis, SYBR Green I
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