地高辛3′末端标记检测效率的分析

Analysis of the Detection Efficiency of Digoxin Labeling at 3′ End

[目的]为核酸杂交试验提供依据。[方法]利用3个不同长度的核酸探针进行3′末端地高辛标记,并对标记效率进行检测,研究探针的长度和浓度及其在尼龙膜上的固定方法对其信号强度的影响。[结果]聚丙烯酰胺凝胶电泳结果表明地高辛标记的探针比未标记探针滞后,说明标记较为成功。检测信号强度随探针摩尔浓度的降低而降低。探针的固定方法对信号强度也有一定的影响。紫外交联和高温烘烤均能使检测信号增强,而使用紫外交联的信号强度较高。[结论]地高辛检测的信号强度与探针的长度、浓度、固定方法以及核酸与膜的结合效率有关。

[Objective] The aim of the research was to provide basis for nucleic acid hybridization test.[Method] Three probes of nucleic acid with different length were used to label at 3′ end with Digoxin and detect its labeling efficiency.The effects of the length and concenstration of probes and its fixation methods on nylon membrane on the signal intensity were studied.[Result] The results of polyacrylamide gel electrophoresis showed that the probes with Digoxin labeling was hysteresis than that without labeling,which indicated that the labeling was more successful.The detected signal intensity was reduced with the reduction of concenstration of probe.The fixation method of probe also had certain effects on the signal intensity.Both ultraviolet cross-linking and high temperature firing could make the detection signal be strengthened.And the signal intensity at ultraviolet cross-linking was higher.[Conclusion] The detected signal intensity of Digoxin was related with the length,concentration,fixation and the comb ined efficacy between nuceleic acid and membrane.