[Objective] This study aimed to explore an effective method for rapid salt- extraction of high-quality genomic DNA from dried seeds of plants. [Method] Seeds of seven varieties of crops were ground into powder. A hund...[Objective] This study aimed to explore an effective method for rapid salt- extraction of high-quality genomic DNA from dried seeds of plants. [Method] Seeds of seven varieties of crops were ground into powder. A hundred milligrams of seed powder was added to extracting solution for high salt-extraction of genomic DNA. The yield and quality of extracted DNA were determined by using ultramicro UV/Vis spectrophotometer detection method, PCR and restriction enzyme digestion. [Result] About 619.67-1 811.21 ng of genomic DNA was extracted from per 100 mg of dried seed powder of seven varieties of conventional crops. A260/A280 ratios of the obtained DNA solution all ranged from 1.87 to 2.07, the purity and quality of PCR were suitable for PCR and restriction enzyme digestion. Clear target bands of specific endogenous gene fragments of seven varieties of crops were amplified by PCR, and the obtained DNA could be fully digested with EcoRV and Hindlll.[Conclusion] This method could be used for rapid extraction of high-quality genomic DNA from dried seeds.展开更多
基金Supported by Project of Common Safety Assessment Technology for Genetically Modified Organisms of the Ministry of Agriculture of PRC (2011ZX08011-006)~~
文摘[Objective] This study aimed to explore an effective method for rapid salt- extraction of high-quality genomic DNA from dried seeds of plants. [Method] Seeds of seven varieties of crops were ground into powder. A hundred milligrams of seed powder was added to extracting solution for high salt-extraction of genomic DNA. The yield and quality of extracted DNA were determined by using ultramicro UV/Vis spectrophotometer detection method, PCR and restriction enzyme digestion. [Result] About 619.67-1 811.21 ng of genomic DNA was extracted from per 100 mg of dried seed powder of seven varieties of conventional crops. A260/A280 ratios of the obtained DNA solution all ranged from 1.87 to 2.07, the purity and quality of PCR were suitable for PCR and restriction enzyme digestion. Clear target bands of specific endogenous gene fragments of seven varieties of crops were amplified by PCR, and the obtained DNA could be fully digested with EcoRV and Hindlll.[Conclusion] This method could be used for rapid extraction of high-quality genomic DNA from dried seeds.
