一种提高基因芯片检测中杂交信号强度的简单方法

Nested coamplification polymerase chain reaction

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。

Objective To develop a simple and reliable method for intensifying the hybridization signals of gene chips. Methods The authors added EDTA and another FAM labeled probe to the normal PCR products, denatured the mixture by heat, and then let the mixture hybridize with the fastened probes on the chip. Results With the use of EDTA and another FAM labeled probe, the hybridization signals increased by 6 times or greater. Conclusion Adding EDTA and another probe to the normal PCR products is a simple and efficient method to intensify the hybridization signal of chips.