党参根组织RNA提取的研究与优化

Study and Optimization of Method for High-quality RNA Extraction from Codonopsis pilosula Root Tissues

目的：建立党参根组织高质量总RNA的提取方法。方法：针对党参根组织多糖含量高的特点，比较了CTAB—LiCl，CTAB—PVP，改良Trizol法3种总RNA的提取方法。结果：CTAB—LiCl法无法提取得到总RNA，CTAB—PVP法提取的总RNA有多糖等杂质污染，改良Trizol法提取的总RNA无DNA及其它杂质污染，党参根组织的产量为120．8μg·g-1，A260／A280在1．8-2．0间，A260／A230大于2．0。结论：改良Trizol法操作简单，提取时间短，提取的RNA可以用于后续分子生物学研究。

Objective： To establish an efficient approach for isolating high -quality total RNA from root of Codonopsis pilosula. Methods ： Extraction of high - quality total RNA from Codonopsis pilosula root was difficult due to high levels of polysaccharides. The methods such as CTAB - LiC1, CTAB - PVP,optimized Trizol were applied to isolate high purity and integrity RNA from Codonopsis pilosula root. Results： Total RNA can not be obtained by CTAB - LiC1. RNA from the CTAB -PVP method was contaminated with polysaccharides, but only by the optimized Trizol method, it extracted high class RNA from root tissue, and there was no obvious contamination with DNA and polysaccharides. The optimized Trizol method isolated RNA from root tissues of Codonopsis pilosula with a yield of 120.8 μg·g-1. The absorbance ratio （ A260 / A280 ） was 1.8 - 2.0. The absorbance ratio （ A260 / A230 ） was above 2.0. Conclusion ： This simple and time - saving system for isolation of RNA was essential to any study related to gene expression.