定量PCR法による遺伝子組換えトウモロコシの定量分析に適用される4種のDNA抽出法の比較検討

概要

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A method based on the real-time polymerase chain reaction (PCR) has been developed as the Japanese official standard method for the quantitative analysis of approved genetically modified (GM) maize by the Ministry of Health, Labour, and Welfare (MHLW) and the Ministry of Agriculture, Forestry, and Fisheries (MAFF). MHLW also defines three methods to extract genomic DNA used as analytes in the real-time PCR method. These three DNA extraction methods include a method using a silica-gel membrane kit (mini method), a silica-based resin type kit (WIZARD method), and the method using a buffer containing cetyltrimethylammmonium bromide (CTAB method). On the other hand, the DNA extraction method using a larger size silica-gel membrane kit (MAXI method), which was employed in collaborative studies to evaluate the performance of the real-time PCR method, has been identified by MAFF for the same purpose. However, the influence of the DNA extraction methods on the GM maize quantitative value using the real-time PCR method has not been demonstrated. Therefore, genomic DNA was extracted from the mixed flour-sample containing a known amount of MON810 and GA21 line, according to four different DNA extraction methods, and analyzed by the real-time PCR method with the aim of examining the influence of the extraction methods on the GM maize quantitative value.
日本食品化学学会の論文
2006-08-31