To develop a simple and fast method for screening genetically modified ingredients from processing by-product and waste,direct quantitative PCR(qPCR)kit-Taqman which omitting multi genomic DNA preparing steps was deve...To develop a simple and fast method for screening genetically modified ingredients from processing by-product and waste,direct quantitative PCR(qPCR)kit-Taqman which omitting multi genomic DNA preparing steps was developed in this study.A total of 18 oil crop processing by-products and wastes including 10 soybean and 8 cotton materials were collected from food processing factories.Compared with 2 commercial direct qPCR kits,conditions of DNA releasing procedure and PCR amplification were optimized.Element screening was performed at the initial step of genetically modified(GM)ingredient testing procedure via direct qPCR.GM event identification was carried out in positive samples by initial screening.Totally 5 screening elements(P–35S,T-NOS,Cp4-epsps,bar and pat)for soybean materials and 6 screening elements(P–35S,T-NOS,NPTII,Cry1Ac,bar and pat)for cotton samples were detected.In GM event identification,MON531 and MON1445 were found in cotton materials.Results were further confirmed by real-time PCR with DNA extraction and purification.The direct qPCR system proposed by this research was convenient for rapid screening and identification of GM ingredients in oil crop primary by-product and waste.展开更多
基金supported by grants from the National Major Special Project of Breeding for Genetically Modified Organisms in China(No.2016ZX08012-005,2016ZX08012-003)the Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences,China.
文摘To develop a simple and fast method for screening genetically modified ingredients from processing by-product and waste,direct quantitative PCR(qPCR)kit-Taqman which omitting multi genomic DNA preparing steps was developed in this study.A total of 18 oil crop processing by-products and wastes including 10 soybean and 8 cotton materials were collected from food processing factories.Compared with 2 commercial direct qPCR kits,conditions of DNA releasing procedure and PCR amplification were optimized.Element screening was performed at the initial step of genetically modified(GM)ingredient testing procedure via direct qPCR.GM event identification was carried out in positive samples by initial screening.Totally 5 screening elements(P–35S,T-NOS,Cp4-epsps,bar and pat)for soybean materials and 6 screening elements(P–35S,T-NOS,NPTII,Cry1Ac,bar and pat)for cotton samples were detected.In GM event identification,MON531 and MON1445 were found in cotton materials.Results were further confirmed by real-time PCR with DNA extraction and purification.The direct qPCR system proposed by this research was convenient for rapid screening and identification of GM ingredients in oil crop primary by-product and waste.
