适于cDNA-AFLP的黄瓜幼叶总RNA快速高效提取方法

A Quick and High Efficient Method for Isolation of RNA from Young Cucumber Leaves

为了快速高效地提取黄瓜幼叶总RNA，满足于cDNA-AFLP实验的要求。对Trizol试剂提取总RNA的方法进行了改进，所提RNA经琼脂糖电泳检测，其28S、18S、5S3条带清晰，完整性好，每0．1g幼叶可获得RNA100～150μg，产率高；经反转录、双链cDNA合成和cDNA-AFLP检测，证实提取的RNA能满足反转录和cDNA-AFLP对RNA质量和数量的要求。

In order to isolate RNA quickly and efficiently from young cucumber leaves for cDNA- AFLP experiment, the method of Trizol isolation of total RNA was modified to efficiently isolate high quality total RNA from young cucumber leaves. Three clear bands of 28S, 18S and 5S could be seen clearly by agarose gel electrophoresis and no degradation was observed. The yield of isolated RNA was high and from 100 μg to 150μg RNA per 0.1 grams of young leaves could be harvested. The results from synthesis of double strand cDNA and cDNA-AFLP indicated that the quality and quantity of the isolated RNA was sufficient for conducting those experiments. This method can be used to isolate RNA from young cucumber leaves for cNDA - AFLP.