Loop-mediated isothermal amplification (LAMP) as a diagnostic tool has been widely used for fish authenticity purpose. An important condition for a successful LAMP amplification is an efficient DNA-extraction procedure. Till now, there has been an overwhelming variety of DNA extraction methods in the literature, yet a comprehensive comparative study for LAMP application is lacking. In this study, five different DNA extraction methods (including Takara, Tiangen, SDS, Tagliavia, and Heating methods) were compared in terms of their ability to extract high yield of pure DNA from Atlantic salmon(Salmo salar) tissue under four different treatments. Moreover, LAMP was employed to evaluate the quality and suitability of the extracted DNA for molecular applications. The results showed that regardless of the extraction methods and sample treatments, the positive LAMP results can always be obtained in agarose gel electrophoresis and naked-eye inspection. While highly significant difference with the Ct values (P<0.01) was observed according to both the extraction methods and sample treatments in real-time fluorescence LAMP assay. The SDS and Tiangen methods, in spite of the lower yield, obtained DNA with higher purity and better sensitivity in LAMP amplification. The results provide a basis for the DNA extraction and detection of Atlantic salmon and the study of different extraction methods suitable for LAMP.
LI Qiuping
,
XU Wenjie
,
CUI Xiaowen
,
CAO Min
,
XIONG Xiaohui
,
XIONG Xiong
. Comparative analysis of different DNA extraction methods for loop-mediated isothermal amplification to identify Atlantic salmon[J]. Food and Fermentation Industries, 2022
, 48(10)
: 240
-246
.
DOI: 10.13995/j.cnki.11-1802/ts.028858
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