cDNA芯片技术杂交效率的优化

目的cDNA芯片杂交影响因素很多,如固定于玻片的DNA片段的浓度、溶解DNA片段的点样液、荧光探针浓度及纯度杂交液、杂交温度、杂交时间和Cy3与Cy5荧光素标记探针等,本实验针对其中重要影响因素进行优化筛选,以提高杂交效率。方法通过逐一改变cDNA杂交技术重要影响因素的实验条件,平行设置多组实验,从中筛选出最佳实验数据。结果与结论重复实验结果表明,杂交效率较高的条件为:浓度约为0.50 μg·μL^-1 DNA片段，溶于1×Micro Spotting Solution(ArrayIt^TM)中，与纯化后的荧光标记在0.5%SDS/10×SSC甲酰胺杂交液，在42℃杂交。

OPTIMIZATION OF HYBRIDIZATION EFFICIENCY IN cDNA CHIP TECHNOLOGY

AIM: There are many factors affecting hybridization in cDNA chip: DNA concentration immobilized on glass surface, spotting solution dissolving DNA, the concentration and purity of fluorescence-labeled probe, hybridization solution, hybridization temperature, hybridization time, Cy3- and Cy5-labeled probes, etc. In order to improve hybridization efficiency, tests were designed to optimize these factors. METHODS: Factors are changed one by one. Optimal values are selected by comparing those of different groups. RESULTS AND CONCLUSION: The efficient hybridization condition is as follows: 0.50 microgram.microL-1 DNA fragments resolved in 1 x Micro Spotting Solution (ArrayIt) hybridize with purified fluorescent probe in 0.5% SDS/10 x SSC at 42 degrees C.