针对转基因玉米的特异基因(GenBank accession number AX342369)设计引物,建立转基因玉米NK603品系特异性基因环介导等温扩增方法。对该方法进行特异性、灵敏度分析。结果表明:LAMP法检测转基因玉米NK603品系成分特异性强、灵敏度高、...针对转基因玉米的特异基因(GenBank accession number AX342369)设计引物,建立转基因玉米NK603品系特异性基因环介导等温扩增方法。对该方法进行特异性、灵敏度分析。结果表明:LAMP法检测转基因玉米NK603品系成分特异性强、灵敏度高、定性检出限为0.1%。该方法快速、简便,可在作物转基因筛查中推广使用。展开更多
In this study, the event-specific real-time fluorescence quantitative PCR method established by Siehuan Academy of Agricultural Sciences was employed to detect the content of flanking fragment （specific fragment） in...In this study, the event-specific real-time fluorescence quantitative PCR method established by Siehuan Academy of Agricultural Sciences was employed to detect the content of flanking fragment （specific fragment） in samples containing 2% genetically modified maize event NK603. The uncertainty of detection results was evaluated based on various uncertainty sources, such as PCR amplification system, data analysis and micropipette. The results showed that A-type uncertainty （ uA ）, B-type uncertainty （ uB ）, combined standard uncertainty （ uC ） and expanded uncertainty （ U95 ） were 0. 000 8,0.1301,0. 001 and 0. 002, respectively; the final detection result was 1.9% ±0.002. Thus, the main uncertainty in detecting flanking fragment of genetically modified maize event NK603 with realtime fluorescence quantitative PCR method was derived from the random effect in the experimental process.展开更多
[ Objective ] The aim of this study was to develop a quantitative PCR detection method for genetically modified maize event NK603, so as to provide ba- sis for quantitative analysis of event NK603. [ Methods ] A quant...[ Objective ] The aim of this study was to develop a quantitative PCR detection method for genetically modified maize event NK603, so as to provide ba- sis for quantitative analysis of event NK603. [ Methods ] A quantitative PCR detection method for genetically modified maize event NK603 was developed using primers and Taqman probe designed according to the flanking sequence of event NK603, which was then adopted to detect the samples containing 2% NK603 stand- ard （with uncertain quantity of 10% ）. [ Results ] The slope of standard curve ranged between -3.6 and -3.1, and the correlation coefficient was higher than 0. 99. The amplification efficiency of this method reached 100.2%, fallen between 90% and 110%. The detected quantity of the experimental sample was 1.9%, closer to the true quantity （2%）. [ Conclusion] This quantitative PCR detection method for genetically modified maize event NK603 is very precise and can be a- dopted in routine testing analysis.展开更多
文摘针对转基因玉米的特异基因(GenBank accession number AX342369)设计引物,建立转基因玉米NK603品系特异性基因环介导等温扩增方法。对该方法进行特异性、灵敏度分析。结果表明:LAMP法检测转基因玉米NK603品系成分特异性强、灵敏度高、定性检出限为0.1%。该方法快速、简便,可在作物转基因筛查中推广使用。
基金Supported by Youth Fund of Sichuan Academy of Agricultural Sciences(2009QNJJ-037)
文摘In this study, the event-specific real-time fluorescence quantitative PCR method established by Siehuan Academy of Agricultural Sciences was employed to detect the content of flanking fragment （specific fragment） in samples containing 2% genetically modified maize event NK603. The uncertainty of detection results was evaluated based on various uncertainty sources, such as PCR amplification system, data analysis and micropipette. The results showed that A-type uncertainty （ uA ）, B-type uncertainty （ uB ）, combined standard uncertainty （ uC ） and expanded uncertainty （ U95 ） were 0. 000 8,0.1301,0. 001 and 0. 002, respectively; the final detection result was 1.9% ±0.002. Thus, the main uncertainty in detecting flanking fragment of genetically modified maize event NK603 with realtime fluorescence quantitative PCR method was derived from the random effect in the experimental process.
基金Supported by Youth Science and Technology Program of Sichuan Academy of Agricultural Science(2009QNJJ-037)Program for Monitoring Invasive Species of Ministry of Agriculture
文摘[ Objective ] The aim of this study was to develop a quantitative PCR detection method for genetically modified maize event NK603, so as to provide ba- sis for quantitative analysis of event NK603. [ Methods ] A quantitative PCR detection method for genetically modified maize event NK603 was developed using primers and Taqman probe designed according to the flanking sequence of event NK603, which was then adopted to detect the samples containing 2% NK603 stand- ard （with uncertain quantity of 10% ）. [ Results ] The slope of standard curve ranged between -3.6 and -3.1, and the correlation coefficient was higher than 0. 99. The amplification efficiency of this method reached 100.2%, fallen between 90% and 110%. The detected quantity of the experimental sample was 1.9%, closer to the true quantity （2%）. [ Conclusion] This quantitative PCR detection method for genetically modified maize event NK603 is very precise and can be a- dopted in routine testing analysis.
