A Modified Method For Total Ribonucleic Acids Extraction From Taxus Chinensis Needles

Taxus chinensis (Pilger) Rehd. is one of the most important medicinal plants. According to the characteristics of the needles, a new RNA extraction method, CTAB-LiCl, was developed. RNA was extracted from young and mature needles of Taxus Chinensis by TRIzol method. The result showed that the quality of RNA extracted from young needles was complete, and that of RNA from mature needles was incomplete. As a comparison, cetyl trimethyl ammonium bromide (CTAB) method, SDS-phenol method and improved CTAB-LiCl method were respectively used for extraction of RNA from mature needles. The comparative result showed that the quality of RNA extracted by CTAB method and SDS-phenol method was incomplete, accompanied with different degrees of contamination of polysaccharides, polyphenols and DNA. The improved CTAB-LiCl method could better extract RNA from mature needles. The ratio of A(260)/A(280) was in the range of 1. 9-2. 0, and the yield of RNA was the highest, which was 1. 37 and 1. 24 times compared with that by CTAB and SDS phenol extraction, respectively. The reverse transcription cDNA was further tested through PCR with the specific primers. The amplified fragments displayed in clear and bright bands in accordance with the expected size. High quality RNA from mature needles has more advantages in molecular biology study of Taxus chinensis.