磷酸甘露糖异构酶基因快速初筛方法的建立
Food Research and Development(2011)
Abstract
建立磷酸甘露糖异构酶(PMI)基因环介导等温扩增快速检测方法。设计了两对内引物和两对外引物,优化后的反应条件为:FIP,BIP浓度为0.8μmol/L,F3,B3浓度为0.2μmol/L,镁离子浓度为2.0 mmol/L,甜菜碱浓度为1.6 mmol/L,反应时间为40 min,反应温度为62.0℃。SYBR Green法检测需2 h,电泳法检测共需2.5 h。与实时荧光和普通PCR比较特异性良好,转基因成分检测限达到0.01%。结果表明该检测方法简单且快速。
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Key words
genetically modified,detection,Loop-mediated Isothermal Amplification,Phosphomannose Isomerase
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