基因芯片的制备研究

Optimization and development of DNA microarray technique

目的 :建立基因芯片的制备技术。 方法 :基因样本准备 ,将带荧光标记的 PCR产物用点样仪点于玻片介质上 ,并经过点样后处理制备成基因芯片 ,用扫描仪测定处理前后芯片上荧光强度的变化 ,计算 DNA固定率。分别设计了 5种玻片 3种固定条件和 6种点样后处理方法的固定率测定。结果 :对芯片制备的各种条件和方法优化实验表明 ,c DNA的氨基修饰 ,点样后的 UV交联和芯片室温干燥处理能有效提高 DNA的固定率。结论 :本研究建立的基因芯片制备技术有较高的效率和可靠的实用性能。

Objective:To compare and optimize conditions of DNA coupling to glass slides for development of DNA microarray. Methods: The process included preparation of DNA samples, spotting and post processing of arrays. DNA microarrays were prepared by spotting fluorescence labeled PCR products of target genes onto specially treated glass slides with robotics. The fluorescent signals before and after treatment were scanned with scanner, and the DNA attachment rate was calculated from the data obtained by ImaGene3.0 software. Results: Several key steps of this technique have been optimized, this study provided a foundation for optimizing conditions DNA attachment in the creation of DNA microarray. Conclusion: Amino modified DNA and UV cross link is useful for anchoring DNA in some distance. Drying the chip in room trmperature after spotting can enhance the DNA combination rate. [