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目的设计一种带有通用序列标签的寡核苷酸探针以应用于寡核苷酸芯片点样的质量控制。方法以HIV寡核苷酸探针作为核心序列,在其一端或两端连接上多种通用序列标签(UST)构建新型探针。然后打印芯片,以Cy5标记的通用引物(Cy5-UP,与UST互补)进行杂交,筛选出5条荧光信号强的探针,重新打印芯片。再分别以不同浓度梯度的、Cy5标记的通用引物(Cy5-UP)和随机引物(Cy5-RP)与芯片杂交,比较两者杂交效率。结果 Cy5-UP的杂交结果显著优于Cy5-RP的杂交结果,特别是在较低浓度时。结论利用Cy5标记的通用引物检测探针上的通用序列标签,可以为芯片点样的质量控制提供一种更有效的方法。
Objective To design an oligonucleotide probe with a universal sequence tag for application to spot-like quality control of oligonucleotide chips. Methods HIV oligonucleotide probes were used as the core sequence, and a variety of universal sequence tags (USTs) were connected to one or both ends to construct novel probes. The chips were then printed and hybridized with Cy5-labeled universal primer (Cy5-UP, complementary to UST) to screen out 5 strong fluorescent probes to reprint the chip. Then, the hybridization efficiency of Cy5-UP and Cy5-RP with different concentration gradient was compared with that of the chip. Results Cy5-UP hybridization results were significantly better than Cy5-RP hybridization results, especially at lower concentrations. Conclusion The universal primer of Cy5-labeled universal primer can provide a more effective method for chip-like quality control.