Comparison of three different methods for total RNA extraction from Fritillaria unibracteata, a rare Chinese medicinal plant

摘要

Efficient RNA isolation is a prerequisite to elucidate the molecular mechanisms of peimine metabolism in the medicinal plant?Fritillaria unibracteata, a rare and famous region drug in Sichuan, China. Three different methods were tested: modified SDS acid phenol, modified CTAB and SDS-LiCl. Compared with the modified CTAB and SDS-LiCl methods, the method of modified SDS acid phenol would obtain initial, abundant and high-quality intact RNA from?F. unibracteata. Meanwhile, the RNA-containing bands are clear and light, and the lightness of 28S rRNA is two times than 18S rRNA. The ratio of A260/A280?is approximately 1.85, and the yield of total RNA is 73 μg per g starting tissue. The specific band of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) gene by the reverse transcription-polymerase chain reaction (RT-PCR) showed that RNA extraction using the modified SDS acid phenol method can provide fundamental materials for further study of genetic modification in this precious Chinese medicinal plant?F. unibracteata.