Evaluation of the RNA extraction methods in different Ginkgo biloba L. tissues

Abstract

Understanding the essential processes of heredity, mechanisms of metabolism, and evolutionary traits of this ginkgo (Ginkgo biloba L.) in terms of molecular biology is necessary. The extraction of high-quality and high-integrity RNA from ginkgo is one of the basic techniques of molecular biology. Therefore, this study aimed to compare the efficacy of the six commonly used RNA extraction methods for detection. TRIzol, modified TRIzol, cetyltrimethylammonium bromide (CTAB), modified CTAB, sodium dodecyl sulfate (SDS), and RNeasy Plant Mini Kit (Qiagen) methods, were evaluated for their capacity to extract high-quality and high-quantity RNA from the embryo and endosperm of ginkgo. The advantages and disadvantages of each method were analyzed to evaluate the isolation process. The optimized modified CTAB method for embryo prevented the precipitation of RNA for a long period and reduced complex separation and precipitation processes. In addition, the modified TRIzol method is more effective in endosperm. Obvious differences in RNA quality and quantity were observed between the embryo and endosperm when the same method is used. Modified TRIzol, modified CTAB, SDS and RNeasy Plant Mini Kit (Qiagen) methods were applied to RNA extraction from other different ginkgo tissues. The modified CTAB is efficient and recommended for the isolation of good-quality total RNA from all tissues with reduced chemical use, costs, and RNA loss. The results of quantitative real-time PCR confirmed that modified CTAB methods could obtain showing low Ct values and high amplification efficiency and enabled the successful isolation of RNA from all tissues.