Comparison of Three DNA Extraction Methods for Feed Products and Four Amplification Methods for the 5 '-Junction Fragment of Roundup Ready Soybean

文献类型： 外文期刊

第一作者： Wang, Xiumin

作者： Wang, Xiumin;Teng, Da;Tian, Fang;Guan, Qingfeng;Wang, Jianhua;Wang, Xiumin;Teng, Da;Tian, Fang;Guan, Qingfeng;Wang, Jianhua

作者机构：

关键词： comparison;genetically modified soybean;DNA extraction;detection;LAMP;HRCA;feed

期刊名称：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY （ 影响因子：5.279； 五年影响因子：5.269 ）

ISSN： 0021-8561

年卷期： 2012 年 60 卷 18 期

页码：

收录情况： SCI

摘要： Three methods of DNA extraction from feed products and four detection methods for the 5'-junction fragment of genetically modified (GM) Roundup Ready soybean (RRS) were compared and evaluated. The DNA extraction methods, including cetyltrimethylammonium bromide (CTAB), sodium dodecyl sulfate (SDS), and guanidine hydrochloride (Kit), were assessed for their yields and purity of DNA, extraction time, and reagent cost. The DNA yields of CTAB, SDS, and Kit were 52-694, 164-1750 and 23-105 ng/mg sample, and their extraction time was 2.5-3,2-2.5, and 1.5-2 h with reagent cost about US dollar 0.24, 0.13, and 1.9 per extraction, respectively. The SDS method was generally well suited to all kinds of feed matrices tested. The limits of detection for the four amplification protocols, including loop-mediated isothermal amplification (LAMP), hyperbranched rolling circle amplification (HRCA), conventional polymerase chain reaction (PCR), and real-time PCR, were 48.5, 4.85, 485, and 9 copies of the pTLH10 plasmid, respectively. The ranked results of the four detection methods were based on multiattribute utility theory as follows (from best to worse): HRCA, LAMP, PCR, and real-time PCR. This comparative evaluation was specifically useful for selection of a highly efficient DNA extraction or amplification method for detecting different GM ingredients.

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