根据外源插入序列与大豆内源基因边界序列,用设计软件Primer Explorer Version 3设计并筛选了转基因大豆GTS 40-3-2品系的环介导等温扩增引物,并进行了反应体系和反应条件的优化,建立了转基因大豆GTS 40-3-2品系特异性环介导等温扩增检...根据外源插入序列与大豆内源基因边界序列,用设计软件Primer Explorer Version 3设计并筛选了转基因大豆GTS 40-3-2品系的环介导等温扩增引物,并进行了反应体系和反应条件的优化,建立了转基因大豆GTS 40-3-2品系特异性环介导等温扩增检测方法。对该方法进行了特异性、灵敏度、稳定性评价,结果显示:该方法能够特异、稳定、灵敏地检测大豆及其制品中的转基因大豆GTS 40-3-2成分,检测低限达到0.5%。此外,对12份实际样品的检测结果表明,该方法与实时荧光PCR方法的检测性能相当,结果吻合率为100%,假阳性率和假阴性率均为0。展开更多
首次以转基因大豆中结构特异性基因为靶基因,建立了特异的微滴式数字QX200 PCR(Droplet digital PCR,ddPCR)检测方法,对其进行准确定量。结果表明:ddPCR对结构特异性基因的检出限为2拷贝/(20μL),检测灵敏度高于国标中qPCR方法。微滴式...首次以转基因大豆中结构特异性基因为靶基因,建立了特异的微滴式数字QX200 PCR(Droplet digital PCR,ddPCR)检测方法,对其进行准确定量。结果表明:ddPCR对结构特异性基因的检出限为2拷贝/(20μL),检测灵敏度高于国标中qPCR方法。微滴式数字PCR方法是一种准确、高灵敏度的基因拷贝数的分析方法,操作简便,可在转基因检测领域广泛应用。展开更多
[ Objective ] This study aimed to investigate the major contributors to the measurement uncertainty in quantitative analysis of genetically modified ingreclients and improve the quality of quantitative detection of ge...[ Objective ] This study aimed to investigate the major contributors to the measurement uncertainty in quantitative analysis of genetically modified ingreclients and improve the quality of quantitative detection of genetically modified components. [ Method] The content of CaMV35S promoter （parameter） in GTS40- 3-2 soybean powder samples was measured to estimate the measurement uncertainty preliminarily. [ Result] Type A uncertainty （uA） ＇ type B uncertainty （uB） and combined standard uncertainty （Uc） were 0.0 004, 0.002 and 0.002, respectively. At a confidence level ofp = 95% and freedom degree of Voff = 3 251, coverage factor k = 1.96, expanded uncertainty U = 0.004. The final measurement result was C = 0.028 ± 0. 004, which was dose to the conventional true value （0.03）. Thus, the measurement uncertainty was relatively small, indicating a high quality of measurement. In this study, uncertainty evaluation indicated that the deviation of micro liquid transfer made the greatest contribution to the measurement uncertainty. [ Cludusion ] The deviation of micro liquid transfer should be reduced to im- prove the quality of measurement.展开更多
Herbicide tolerant plants such as Roundup-Ready soybean contain residues of glyphosate herbicide. These residues are considered safe and previous animal-feeding-studies have failed to find negative effects related to ...Herbicide tolerant plants such as Roundup-Ready soybean contain residues of glyphosate herbicide. These residues are considered safe and previous animal-feeding-studies have failed to find negative effects related to such chemical residues. The present study tests 8 experimental soy- meal diets as feed in groups (each containing 20 individuals) of test-animals (D. magna). The diets have different levels of glyphosate residues and we show that animal growth, reproductive maturity and number of offspring are correlated with these chemicals. The tested soybeans are from ordinary agriculture in Iowa USA and the residues are below the regulatory limits. Despite this, clear negative effects are seen in life-long feeding. The work enhances the need for including analysis of herbicide residues in future assessment of GMO.展开更多
文摘根据外源插入序列与大豆内源基因边界序列,用设计软件Primer Explorer Version 3设计并筛选了转基因大豆GTS 40-3-2品系的环介导等温扩增引物,并进行了反应体系和反应条件的优化,建立了转基因大豆GTS 40-3-2品系特异性环介导等温扩增检测方法。对该方法进行了特异性、灵敏度、稳定性评价,结果显示:该方法能够特异、稳定、灵敏地检测大豆及其制品中的转基因大豆GTS 40-3-2成分,检测低限达到0.5%。此外,对12份实际样品的检测结果表明,该方法与实时荧光PCR方法的检测性能相当,结果吻合率为100%,假阳性率和假阴性率均为0。
文摘首次以转基因大豆中结构特异性基因为靶基因,建立了特异的微滴式数字QX200 PCR(Droplet digital PCR,ddPCR)检测方法,对其进行准确定量。结果表明:ddPCR对结构特异性基因的检出限为2拷贝/(20μL),检测灵敏度高于国标中qPCR方法。微滴式数字PCR方法是一种准确、高灵敏度的基因拷贝数的分析方法,操作简便,可在转基因检测领域广泛应用。
基金Supported by Project of Standardized Technology System of Sichuan Bureau of Quality and Technical Supervision(ZYBZ2013-39)
文摘[ Objective ] This study aimed to investigate the major contributors to the measurement uncertainty in quantitative analysis of genetically modified ingreclients and improve the quality of quantitative detection of genetically modified components. [ Method] The content of CaMV35S promoter （parameter） in GTS40- 3-2 soybean powder samples was measured to estimate the measurement uncertainty preliminarily. [ Result] Type A uncertainty （uA） ＇ type B uncertainty （uB） and combined standard uncertainty （Uc） were 0.0 004, 0.002 and 0.002, respectively. At a confidence level ofp = 95% and freedom degree of Voff = 3 251, coverage factor k = 1.96, expanded uncertainty U = 0.004. The final measurement result was C = 0.028 ± 0. 004, which was dose to the conventional true value （0.03）. Thus, the measurement uncertainty was relatively small, indicating a high quality of measurement. In this study, uncertainty evaluation indicated that the deviation of micro liquid transfer made the greatest contribution to the measurement uncertainty. [ Cludusion ] The deviation of micro liquid transfer should be reduced to im- prove the quality of measurement.
文摘Herbicide tolerant plants such as Roundup-Ready soybean contain residues of glyphosate herbicide. These residues are considered safe and previous animal-feeding-studies have failed to find negative effects related to such chemical residues. The present study tests 8 experimental soy- meal diets as feed in groups (each containing 20 individuals) of test-animals (D. magna). The diets have different levels of glyphosate residues and we show that animal growth, reproductive maturity and number of offspring are correlated with these chemicals. The tested soybeans are from ordinary agriculture in Iowa USA and the residues are below the regulatory limits. Despite this, clear negative effects are seen in life-long feeding. The work enhances the need for including analysis of herbicide residues in future assessment of GMO.
