Extraction of RNA from Recalcitrant Tree Species Paulownia elongata

摘要

Isolation of pure RNA is the basic requisite for most molecular biology work. Plants contain polyphenols and polysaccharides, which can interfere with isolation of pure RNA from them. Especially hardwood tree species like Paulownia elongata have surplus amount of RNA-binding alkaloids, proteins and secondary metabolites that can further complicate the process of RNA extraction. Paulownia elongata is a fast-growing tree species which is known for its role in environmental adaptability and biofuel research. Here we describe an economical, efficient and time-saving method (2 h) to extract RNA from leaf tissues of the tree Paulownia elongata. Lack of DNA contamination and good RNA integrity were confirmed using RNA Gel electrophoresis. The purity of RNA was confirmed using Nanodrop spectrophotometer that revealed an A260:A280 ratio of about 2.0. The purified RNA was successfully used in the downstream applications such as RT-PCR (Reverse Transcription PCR) and qPCR (quantitative PCR). This method could be used for RNA extraction from several other recalcitrant tree species.