以纤维素酯膜作为基底材料进行生物芯片杂交条件的优化

Optimization of Condition for Biochip Hybridization Using Cellulose Ester Membrane as A Substrate

目的优化以纤维素酯膜作为基底材料进行生物芯片杂交的条件。方法分别用不同厂家、不同孔径的基底膜进行杂交,筛选最佳基底膜;以不同长度探针片段、不同核酸标记方式进行杂交,筛选最佳探针长度和靶基因标记方法;分别在30、40和50℃杂交0.5、1.0和1.5h,比较芯片杂交效果。结果美国Millipore公司的0.45μm混合纤维素酯膜为最佳基底膜;Biotin-16-dUTP探针标记方法和在40℃杂交1.0h为最佳杂交条件。结论优化了以纤维素酯膜作为基底材料进行生物芯片杂交的条件。

Objective To optimize the condition for biochip hybridization using cellulose ester membrane as a substrate. Methods Biochip hybridization tests were performed by using the cellulose ester membranes, at various apertures, produced by different manufacturers, based on which the length of probe and the method for labeling of target gene were optimized. The tests were further performed by using the optimal cellulose ester membrane at 30, 40 and 50℃ to optimize the temperature, then at the optimal temperature for 0. 5, 1. 0 and 1. 5 h to optimize the time for biochip hybridization. Results By using the optimal cellulose ester membrane （0. 45 m, Millipore）, the condition for biochip hybridization was optimized as labeling with Biotin-16-dUTP probe at 40℃ for 1.0 h. Conclusion The condition for biochip hybridization using cellulose ester membrane as a substrate was optimized.