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五种不同组织结构特点昆虫的总RNA提取效率比较
Comparison of the effectiveness of the Trizol method versus an RNA extraction kit for extracting total RNA from insect species with different tissue characteristics
尹志勇;郭建军;陈绪美;郑玉琳;田 莹;檀 军
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DOI:10.7679/j.issn.2095-1353.2020.102
作者单位:贵州大学昆虫研究所,贵州山地农业病虫害重点实验室 ; 遵义医科大学,组织学与胚胎学教研室
中文关键词:昆虫总RNA;提取方法;提取效率;紫外分光光度;琼脂糖凝胶电泳
英文关键词:insect RNA; extraction method; extraction efficiency; ultraviolet spectrophotometer; agarose gel electrophoresis
中文摘要:
【目的】 比较RNAeasy抽提试剂盒法和Trizol提取法对不同种昆虫总RNA提取的效率,为不同种昆虫总RNA提取方法选择提供参考。【方法】 采用RNAeasy抽提试剂盒法和Trizol提取法分别对九香虫Aspongopus chinensis Dallas、白背飞虱Sogatella furcifera Horváth、中华蜜蜂Apis cerana cerana、黄蜻Pantala flavescens和蒿金叶甲Chrysolina aurichalcea 进行总RNA提取,通过紫外分光光度计测定总RNA吸光度(OD)值,利用琼脂糖凝胶电泳检测等方式对RNA质量进行评估。【结果】 2种提取方法提取不同昆虫总RNA中,Trizol法提取白背飞虱(同翅目)、蒿金叶甲(鞘翅目)和九香虫(半翅目)的总RNA浓度较高,但纯度较低。电泳结果显示总RNA有降解,完整性较差,RNAeasy抽提试剂盒法提取黄蜻(蜻蜓目)和中华蜜蜂(膜翅目)的浓度较低,纯度较高,电泳结果显示完整性较好。【结论】 Trizol法更适合提取小型或几丁质含量高的昆虫,RNA抽提试剂盒法更适合几丁质含量较少、腹软且有大量体液的昆虫总RNA提取。
英文摘要:
[Objectives]  The relative efficiency of using a RNA extraction kit to extract total RNA from different insects was compared to that of the Trizol method. [Methods]  An RNA extraction kit and Trizol were used to extract total RNA from five insects; Aspongopus chinensis Dallas, Sogatella furcifera Horváth, Apis cerana cerana, Pantala flavescens and Chrysolina aurichalcea. The quality and the OD value of total RNA obtained were measured with a UV spectrophotometer and agarose gel electrophoresis. [Results]  Although a higher concentration of total RNA was extracted with the Trizol method from three species, S. furcifera, C. aurichalcea, and A. chinensis, it had lower purity and electrophoresis results indicated that it was degraded with poor integrity. Conversely, although the concentration of total RNA obtained with an RNA extraction kit from P. flavescens (Odonata) and A. cerana cerana (Hymenoptera) was relatively low, it was relatively pure and had better integrity. [Conclusion]  The trizol method is more suitable for extracting total RNA from small insects or those with high chitin content, whereas an RNA extraction kit is better for insects with low chitin content, a soft abdomen and more body fluid. 
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