Optimized condition of genomic DNA extraction and PCR methods for GMO detection in potato

摘要

To compare the quality of genomic DNA extracted from potato for PCR detection, four different methods, such as silica-based membrane method, silica-coated bead method, STE solution treatment, and CTAB-phenol/chloroform method, were evaluated. Also,to remove an excessive carbohydrate from the potato, alpha- and beta-amylase were used individually and in combination. When used both silica-based membrane method and silica-coated bead method combined with enzymes, the genomic DNAs were extracted fromthe raw potato with high purity for PCR. However, the silica-coated bead method combined with enzyme treatment was the most efficient for extraction of the genomic DNA from the frozen fried potatoes. When applied with STE solution, the highly purified DNAwas extracted from the raw potatoes without enzyme treatment in adequate yield for PCR. In cases of processed potatoes, such as frozen-fried potato and fabricated potato chips, CTAB-phenol/chloroform method is mostly feasible for DNA extraction and PCRefficacy at high sensitivity. As the results of PCR amplification, 216bp of PCR product was detected on 2 percent agarose gel electrophoresis, but any amplicons derived from New leaf and New leaf Y gene was not detected in any sample.