基因芯片在差异表达研究中测定影响因素的初步探讨

Analysis of the experimental influences on the procedure of cDNA microarray

目的 :通过对细胞凋亡及信号传导基因芯片在差异表达研究中测定影响因素的分析。方法 :提取小鼠肝脏组织总 RNA,经反转录获得具有荧光标记的 c DNA探针 ,再将探针与芯片杂交 ,洗涤干燥后 ,扫描芯片获取结果。结果 :获得高纯度、高质量的组织总 RNA,检测逆转录反应的效率 ,观察到较理想的扫描芯片图像。结论 :高质量的 RNA及高效率的逆转录反应 。

Objective:To analyze the experimental influences on the procedures of cDNA microarray designed for different expression of apoptosis and signal transduction and to establish and optimize the method for further clinical application.Methods:Total RNA of mouse liver tissue was extracted, cDNA probes were labelled with fluorescence by means of reverse transcription,hybridized to cDNA microarray,and then washed and desiccated.Hybridization signals were scanned and recorded.Results:High quality total RNA was acquired;reaction efficiency of reverse transcription was evaluated;hybridization signals of DNA microarray was scanned and detected clearly.Conclusions:High quality total RNA and reaction efficiency of reverse transcription are crucial to cDNA microarray technology.