An efficient method for extraction of RNA from rice leaves at different ages using benzyl chloride
An efficient method for extraction of RNA from rice leaves at different ages using benzyl chloride
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DOI：
10.1093/jexbot/52.360.1575
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发表时间：
2001-07-01
影响因子：
6.9
通讯作者：
Mae, T
中科院分区：
文献类型：
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作者：
Suzuki, Y;Makino, A;Mae, T
用酸性硫氰酸胍-水饱和的酚-氯仿混合物提取RNA的常规方法，在不同年龄的水稻叶片中，可提取的RNA相对于总RNA（可提取的RNA+残余RNA）的提取效率为54- 69%。改进后的新方法，用氯化苄代替水饱和苯酚，再用少量石英砂进一步浸渍，效率提高到81- 95%。当用改进的方法获得的RNA组分进行琼脂糖凝胶电泳时，检测到25 S和17 S rRNA的完整条带。用水稻rbcS的DNA探针在印迹膜上只观察到一条带。这些结果表明，改进的氯化苄提取RNA方法可用于水稻硬叶等植物组织RNA的定量和定性分析。
With a conventional method of RNA extraction using an acid guanidinium thiocyanate-water-saturated phenol-chloroform mixture, extraction efficiency of extractable RNA to total RNA (extractable RNA+ residual RNA) in rice leaves at various ages was 54-69%. With a new method, an improvement of the above, using benzyl chloride instead of water-saturated phenol together with further maceration with a small amount of quartz sand, the efficiency was increased to 81-95%. When RNA fractions obtained with the improved method were subjected to agarose gel electrophoresis, intact bands of 25 S and 17 S rRNAs were detected. With a DNA probe for rice rbcS, only a single band was observed on the blotted membrane. These results indicate that the improved extraction method of RNA with benzyl chloride is useful for quantitative and qualitative analysis of RNA in plant tissues such as stiff leaves of rice.