南方红豆杉不同组织总RNA提取方法研究

Study on extraction methods of total RNA from different tissues of Taxus mairei(Lemee et LevI.) S. Y. Huex Liu

目的:从南方红豆杉不同组织中提取高质量的总RNA。方法:以南方红豆杉幼叶、嫩茎和种子为材料,采用Trizol法、CTAB法、热酚法、改良热酚法和天根RNAprep Pure多糖多酚植物总RNA提取试剂盒法5种方法进行比较分析。结果:Trizol法基本未提取到南方红豆杉不同组织的RNA;CTAB法提取的RNA浓度较高,但OD_(260)/OD_(280)大于2.1,RNA降解严重;热酚法提取的RNA浓度较高,OD_(260)/OD_(280)均小于1.8,蛋白质和酚类物质等残留严重,质量较低;改良热酚法和试剂盒法28S rRNA、18S rRNA和5S rRNA条带清晰,完整性好,OD_(260)/OD_(280)均在1.8~2.1范围内,纯度高。结论:Trizol法、CTAB法和热酚法均不适用于南方红豆杉总RNA提取,而改良热酚法和试剂盒法可提取到质量较好的南方红豆杉总RNA。

Objective： To extract high quality total RNA from different tissues of Taxus mairei （Lemee et Levi.） S. Y. Huex Liu. Methods： The young leaves, tender stems, and seeds of Taxus mairei （Lemee et Levi.） S. Y. Huex Liu were used as materials to extract the total RNA. The five methods, such as Trizol method, CTAB method, hot phenol method, modified hot phenol method and kit method （RNAprep Pure polysaccharide polyphenol plant RNA extraction ） were used to compared and analyzed. Results： RNA from different tissues of Taxus mairei （Lemee et Levi.） S. Y. Huex Liu was not extracted by Trizol method. The RNA concentration extracted by CTAB method was high, but OD260/OD280 was higher than 2.1, and the RNA degradation was serious. The RNA concentration extracted by hot phenol method was high, and the OD260,/OD280 was lower than 1.8, but the protein and phenolic substances were seriously residual and of low quality. The bands of 28s rRNA, 18s rRNA and 5s rRNA were clear and had good integrity, which were extracted by modified hot phenol method and kit method, and their OD260/ OD280 were both within the range of 1.8~2.1 with high purity. Conclusion： The trizol method and CTAB method are not suitable for the extraction of total RNA from Taxus mairei （Lemee et Levl.） S. Y. Huex Liu, while the modified hot phenol method and kit method can extract better RNA.