摘要
目的:确定高分辨率熔解曲线分析方法可用于PCR条件优化。方法:对家蝇抗性基因cyp6d1和kdr进行温度梯度PCR,其产物分别使用高分辨率熔解曲线分析和溴化乙锭琼脂糖凝胶电泳方法判定最佳退火温度,并将结果进行比较分析。结果:根据高分辨率熔解曲线判定cyp6d1和kdr基因的最佳退火温度分别为65℃和63.8℃,与根据溴化乙锭琼脂糖凝胶电泳判定的退火温度相同。结论:可用高分辨率熔解曲线判断PCR最佳退火温度。
Objective：To provide evidence for using high resolution melting analysis in optimization of PCR condition.Methods： Temperature gradient PCR was performed on two housefly deltamethrin resistance-associated genes,cyp6d1 and kdr.The PCR products were performed high resolution melting analysis and agarose gel electrophoresis with ethidium bromide.The optimal annealing temperature were detected according to the melting curve and fluorescence intensity respectively.Results： According to the melting curve,the optimal annealing temperature for cyp6d1 and kdr were 65℃ and 63.8℃ respectively,which were consistent with the results detected from fluorescence intensity.Conclusion： High resolution melting analysis could be used to optimize PCR condition.
出处
《中国卫生检验杂志》
北大核心
2012年第11期2539-2540,2543,共3页
Chinese Journal of Health Laboratory Technology
基金
浙江省卫生厅科研项目(2009A176)
关键词
高分辨率熔解曲线
PCR条件优化
High resolution melting analysis
Optimization of PCR condition
