摘要
【目的】探索牡丹根系RNA提取的最佳方法。【方法】以牡丹(Paeonia suffruticosa)'凤丹'5a生实生苗的根系为材料,首次比较研究了改良Trizol法、改良CTAB法、改良异硫氰酸胍法及Column Plant RNAout试剂盒法提取RNA的效果。【结果】改良Trizol法及改良异硫氰酸胍法均不能从牡丹根系中提取出高质量RNA。而改良CTAB法及试剂盒法提取RNA的电泳结果可见清晰完整的28S rRNA1、8S rRNA及5S rRNA条带,其中28S rRNA与18S rRNA的亮度比约为2∶1;浓度及纯度检测表明这2种方法提取的RNA纯度较高,且改良CTAB法提取的RNA浓度(269.50μg.g-1)是试剂盒法(82.14μg.g-1)的3倍。【结论】结合RNA的提取质量和成本,改良CTAB法更适宜于牡丹根系RNA的提取。
The roots five-year-old seedlings of of Paeonia suffruticosa ＇Fengdan＇ were used to extract total RNA.Four methods were compared to select optimal one： Trizol method,modified guanidinium thiocyanae method,modified CTAB method,and Column Plant RNAout kit method.The results showed that modified Trizol method and modified guanidinium thiocyanae method were not suitable for extracting the high-quality RNA from the root of P.suffruticosa.The complete 28S rRNA,18S rRNA and 5S rRNA bands could be seen clearly on the electrophoretogram of total root RNA extracted by modified CTAB method and Column Plant RNAout kit method,the brightness ratio of 28S rRNA and 18S rRNA were about 2∶1;detection for concentration and purity showed that total RNA extracted by these two methods were purer.But the concentration of total RNA extracted by the modified CTAB method was as three times as that of the kit method.Considering total RNA purity and reagents cost,the modified CTAB method was more suitable for extracting total RNA from the root of P.suffruticosa.
出处
《西北林学院学报》
CSCD
北大核心
2010年第5期60-63,共4页
Journal of Northwest Forestry University
基金
河南省高校创新人才培养工程(豫教高[2005]126号)
关键词
牡丹
根系
RNA
提取
改良CTAB法
Paeonia suffruticosa
root
RNA
extraction
modified CTAB method
