水稻整体原位杂交实验技术方法的改进

以检测水稻SUPERWOMAN1(SPW1)/OsMADS16基因的表达模式为例,通过对杂交探针的制备、材料的固定及解离通透、显色等方面进行优化,获得背景值低、特异性高的SPW1/OsMADS16基因特异性表达结果.建立一套针对水稻的整体原位杂交技术,方法简单,费用低廉,可在离心管中多种材料同时进行.主要实验步骤包括探...

Improvement of Experimental Techniques of Whole Mount in situ Hybridization Technology in Rice

Taking rice SUPERWOMAN1 (SPW1)/OsMADS16 gene as an example, by optimizing the preparation of hybridization probes, material immobilization, dissociation, permeabilization and color development, the SPW1/OsMADS16 gene expression patterns with high specificity but low backgrounds were obtained. The procedure of whole mount in situ hybridization (WISH) technology for rice is simple and inexpensive, and it can be performed simultaneously with several species in the centrifuge tube. The main experimental steps include probe preparation, material immobilization and dissociation, hybridization reaction, post-hybridization treatment and detection and color development. The proposed method provides a technological basement for a protentional high throughput detection of spatiotemporal gene expression patterns and functional analysis in rice.