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目的:制备高质量的丹参根部RNA与均一化的丹参根部全长cDNA。方法:利用改良的TRIzol方法,从丹参根部提取得到高质量的总RNA,利用SMART技术合成丹参根部全长cDNA,并应用DSN酶处理的方法对丹参根部全长cDNA进行均一化。结果:电泳检测结果表明,利用改良的TRIzol方法提取的总RNA具有较好的完整性,所获得的丹参根部全长cDNA均一化效果良好。结论:本研究表明,改良的TRIzol方法适用于丹参根部组织的RNA提取。本研究中制备的丹参根部全长cDNA,为后续丹参功能基因的克隆及丹参RNA干涉突变体库的构建提供了材料。
OBJECTIVE: To prepare high quality Radix Salviae miltiorrhizae root RNA and homogenized Radix Salviae miltiorrhizae root full length cDNA. Methods: The improved TRIzol method was used to extract high quality total RNA from root of Salvia miltiorrhiza. SMART was used to synthesize full length cDNA of root of Salvia miltiorrhiza. DSN enzyme treatment was used to homogenize the full length cDNA of root of Salvia miltiorrhiza. Results: The results of electrophoresis showed that the total RNA extracted by the improved TRIzol method had good integrity and the homogeneity of full-length cDNA of Salvia miltiorrhiza root obtained was good. Conclusion: This study shows that the modified TRIzol method is suitable for RNA extraction of Salvia miltiorrhiza roots. The full-length cDNA of Radix Salviae Miltiorrhizae prepared in this study provided the material for the cloning of the functional gene of Salvia miltiorrhiza and the construction of Salvia miltiorrhiza RNA interference mutant library.