昆虫总RNA提取及实时荧光定量PCR方法探讨

The application of RNA extraction and realtime fluorescent quantitative PCR in insect

董小林

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作者单位：华南农业大学昆虫毒理研究室广州510642

中文关键词：昆虫，RNA提取，完整性， RTqPCR

英文关键词：insect，RNA extraction，integrality，realtime fluorescent quantitative PCR

中文摘要：

本文以褐飞虱Nilaparvata lugens (Stl)为实例介绍了利用Trizol略法作改进用于提取昆虫总RNA的方法，抽提的总RNA利用甲醛变性凝胶电泳、NanoDrop核酸蛋白仪和Agilent 2100 Bioanalyzer分析表明其质量高和完整性好。同时讨论了应用实时荧光定量PCR (RTqPCR) 进行昆虫时空表达研究中选择参考基因应注意的问题，为相关研究提供参考。

 

英文摘要：Using the brown planthopper as an example, we introduce a modified protocol of RNA extraction using the Trizol reagent method. Formaldehyde modified gel electrophoresis, NanoDrop (Thermo) and Agilent 2100 Bioanalyzer detection indicated that the RNA extracted by this method had high quality and perfect integrality. Several issues in the use of real\|time fluorescent quantitative PCR (RT\|qPCR) technique on insects became apparent in the expression pattern analysis. These results may provide a reference for the application of this technique in entomology.

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