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目的 改进 PCR产物与固定探针杂交效率 ,提高信噪比 ,以提高基因芯片检测的灵敏度。方法 以淋球菌隐蔽型质粒 p JD1的基因芯片检测为例 ,在荧光素标记引物的 PCR产物中加入不同浓度的Taq酶抑制剂 EDTA及位于固定探针所在位点的两翼的荧光素标记探针 ,一起变性后再进行常规杂交、冲洗。结果 加入 EDTA和荧光素标记探针使基因芯片检测的荧光强度能提高 6倍以上。结论 加入EDTA和荧光素标记探针是一种提高基因芯片检测的效率的简单而有效的方法
Objective To improve the hybridization efficiency between PCR products and immobilized probes and improve the signal-to-noise ratio to improve the sensitivity of gene chip detection. Methods Gene chip detection of neisseria gonorrhoeae cryptic plasmid p JD1 was taken as an example. The PCR product of fluorescein-labeled primer was added with different concentration of Taq enzyme inhibitor EDTA and fluorescein-labeled probe located on both wings of the immobilized probe , After denaturation with conventional hybridization, washing. Results The addition of EDTA and fluorescein-labeled probes increased the fluorescence intensity of gene chips by more than 6-fold. Conclusion The addition of EDTA and fluorescein-labeled probes is a simple and effective method to increase the efficiency of microarray detection