【摘要】 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。更多还原

【Abstract】 A loop mediated isothermal amplification(LAMP) assay was developed for the rapid detection of gene PMI.A set of sepcific primers,two inner primers and outer primers were designed.The concentrations: FIP and BIP,both 0.8 μmol/L,F3 and B3,both 0.2 μmol/L,Mg2+,2 mmol/L,Betaine,1.6 mmol/L.The reaction time and temperatures were optimized for 40 min at 62.0 ℃.SYBR Green I detection method can be finished in 2 h.Electrophoresis detection method can be finished in 2.5 h.The LAMP assay has a higher specificity compared with Real Time PCR and conventional PCR.The detection limit of GM elements reaches 0.01 %.The results indicated the suitability and simplicity of the test as a rapid method for PMI specific detection.更多还原