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水稻RNA原位杂交体系的优化

DOI: 10.7685/j.issn.1000-2030.2012.02.003, PP. 15-20

Keywords: 原位杂交,水稻,固定液,消化时间,杂交液

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Abstract:

为了能准确、快捷地追踪目标基因在水稻中的表达,并提供在不同植物中基因表达的研究基础,优化了水稻RNA原位杂交体系。以卡诺固定液处理材料,原位杂交结果显示细胞边界清楚,标本背景清晰,检测信号强。设置了0、15、25、35和45min5个消化时间梯度,消化时间是25min的试验结果显示杂交信号分布于整个受检表面,信号可检测性强,并在不同组织结构中信号强弱不同。以根尖为材料,杂交液A的检测信号符合分布规律,杂交液B验证反义探针检测信号较弱,杂交液C验证反义探针检测信号进一步减弱,并伴随着验证正义探针信号增强。以叶片为试验材料,杂交液A和B的原位杂交结果差异不大,而杂交液C验证反义探针信号减弱,验证正义探针信号增强。结论水稻RNA原位杂交体系的固定液为卡诺固定液,最佳消化时间为25min,杂交液为杂交液A。

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