Comparison of Three DNA Extraction Methods for Feed Products and Four Amplification Methods for the 5鈥?Junction Fragment of Roundup Ready Soybean
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- 作者：Xiumin Wang ; Da Teng ; Fang Tian ; Qingfeng Guan ; Jianhua Wang
- 刊名：Journal of Agricultural and Food Chemistry
- 出版年：2012
- 出版时间：May 9, 2012
- 年：2012
- 卷：60
- 期：18
- 页码：4586-4595
- 全文大小：326K
- 年卷期：v.60,no.18(May 9, 2012)
- ISSN：1520-5118
文摘
Three methods of DNA extraction from feed products and four detection methods for the 5鈥?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鈥?94, 164鈥?750 and 23鈥?05 ng/mg sample, and their extraction time was 2.5鈥?, 2鈥?.5, and 1.5鈥? 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.