An easy and robust method for the isolation of high quality RNA from coconut tissues

ABSTRACT

BACKGROUND: Coconut tissues consist of a complex network of polysaccharides, proteins, polyphenols, and lipids that can bind to nucleic acids and pose difficulty in isolation. Certainly, a vigorous method is required to isolate high quality and quantity of RNA from such tissues for the purpose of downstream experiments. In this paper, we discuss a newly developed method for the Isolation of RNA from Complex Matrices (IRCM) method from coconut tissues. RESULTS: The method is robust, cheap, and efficient for the extraction of quality RNA in high quantities from the solid endosperm of stored and fresh coconut (150 µg/g FW with A260/280 = 1.89 and 247.5 µg/g FW with A260/280 = 1.91), coconut apple (263.8 µg/g FW with A260/280 = 1.97), and coconut bud (1052.5 µg/g FW with A260/280 = 2.00). The other well established methods, such as Method of RNA Isolation from Palm (MRIP), Cetyl Trimethyl Ammonium Bromide (CTAB), TRIZOL, and RNA plant kit failed to isolate quality RNA in appreciable quantities from the coconut tissues. Furthermore, the resultant RNA performed well in the downstream experiment, that is, RT-PCR for the production and amplification of cDNA. CONCLUSIONS: From the study, we concluded that the present method will play a vital role in the extraction of high quality RNA from complex matrices in a short time.