采用试剂盒法、热酚法和LiCI沉淀法3种方法提取葡萄叶片总RNA,通过琼脂糖凝胶电泳和Agilent2100计检测总RNA的完整性和提取纯度。对红地球葡萄叶片总RNA提取方法做比较,以得到完整的、高质量的葡萄RNA。结果表明:LiCI沉淀法提取的RNA收率高,完整性好,OD260/0D280均在1.80~2.00,纯度高,28S、18S条带清晰完整。可以满足后续分子生物学的研究。
Three extraction methods of RNeasy Mini Kit, hot phenol method and LiC1 precipitation were used to extract total RNA from grape leaves,and their integrity and purity were detected by via agarose gel electrophoresis and Agilent 2100 analyser for a comparative study,in order to compare three RNA extraction methods and find out the suitable one for isolating intact and high quality total RNA from grape leaves (Vitis vinifera L. cv. Red Globe). The results showed that among the three extraction methods, LiCl precipitation method demonstrated higher yield and better integrity of total extracted RNA,with a OD260/OD280 ratio between 1.8-2. 0 and clear 28S and 18S bands in electrophoresis pattern. It could meet the need in molecular biological studies.