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以暗紫贝母(Fritillaria unibracteata Kisao et K.C.Hisa)新鲜鳞茎为试材,选取3种有效提取富含油脂、多糖和酚类植物材料总RNA的方法,比较提取总RNA的效果。结果表明,改良SDS酸酚法能有效去除大部分多糖和DNA,RNA条带清晰、亮度好,28S rRNA的亮度约为18S rRNA的2倍,A260nm/A280nm为1.85,A260nm/A230nm为2.06,得率为73μg/g。提取的总RNA经RT-PCR获得了3-羟基-3-甲基戊二酰辅酶A还原酶(3-hydroxy-3-methylglutaryl-CoA reductase,HMGR)基因的特异性条带,说明改良SDS酸酚法从川贝母中提取的RNA质量好、产率高、完整性强,完全适合于进一步的分子生物学研究。
Using fresh bulbs of Fritillaria unibracteata Kisao et K.C.Hisa as test material, three methods for extracting total RNA from plant material rich in oil, polysaccharides and phenols were selected to compare the effect of total RNA extraction. The results showed that the modified SDS acid phenol method can effectively remove most of the polysaccharides and DNA, the RNA bands clear, good brightness, 28S rRNA brightness of about 18S rRNA 2 times, A260nm / A280nm 1.85, A260nm / A230nm 2.06, The rate was 73 μg / g. The specific bands of the 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) gene were obtained by RT-PCR from the extracted total RNA, indicating that the modified SDS acid Phenol extracted from Fritillaria cirrhosa RNA quality, high yield, integrity, fully suitable for further molecular biology.