Comparison of Several Total RNA Extraction Methods from Flowers of Dendrobium officinale

Abstract:

In order to screen total RNA extraction method from flowers of Dendrobium officinale, eight methods were compared, including improved CTAB-LiCl method (M1), improved CTAB-isopropanol method (M2), improved SDS-LiCl method (M3), improved SDS-isopropanol method (M4), Quick RNA Isolation Kit (Huayueyang, China; M5), Column Plant RNAout 2.0 Kit (Tiandz, China; M6), RNAprep Pure Plant Plus Kit (Tiangen, China; M7) and Plant Total RNA Extraction Kit (Biospin, China; M8). The results showed that the integrity of total RNA extracted using M4 and M5 was good, with distinct electrophoresis bands of 28S and 18S rRNA. The spectrophotometric values of A_{260 nm}/A_{280 nm} ranged from 1.8 to 2.0, and the A_{260 nm}/A_{230 nm} value was more than 2.0. The yields of total RNA extracted using M4 and M5 were (159.45±1.45) and (170.84±3.53) μg/g, respectively. Additionally, the total RNA of D. huoshanense, D. nobile, D. chrysotoxum and D. loddigesii extracted by M4 and M5 was verified to meet the quality requirements on integrity, purity and concentration. The total RNA of D. officinale extracted by M4 and M5 was used as template, and the amplified Actin gene showed a single band, consisting with the prediction. Therefore, M4 and M5 were effective methods to extract total RNA from flowers of Dendrobium plants, especially D. officinale.

Key words:Dendrobium officinale;RNA;Improved SDS-isopropanol method;Extraction method