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

Abstract

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.   Key words: Fritillaria unibracteata, RNA extraction, modified SDS acid phenol.