【摘要】 建立耐高温α-淀粉酶Amy797E基因环介导等温扩增快速检测方法。LAMP引物,优化反应条件,FIP,BIP浓度为0.8μmol/L,F3,B3浓度为0.2μmol/L,镁离子浓度为2 mmol/L,甜菜碱浓度为1.4 mmol/L,反应时间为40 min,反应温度为62℃,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%。更多还原

【Abstract】 In this study,a loop mediated isothermal amplification(LAMP) assay was developed for the rapid detection of gene PMI.A set of sepcific primers,two primers,two inner and outer primers and a loop primer,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,1.4 mmol/L.The reaction time and temperatures were optimized for 40 min at 62 ℃.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