An optimized RNA extraction protocol for polyphenol and polysaccharide rich banana tissues

Isolating high-quality RNA from plant tissues rich in polyphenols and polysaccharides presents a significant challenge in molecular research. This study details a robust and reproducible method for extracting total RNA from high-polyphenol tissues of bananas. The protocol begins with 0.5 g of flash-frozen tissue ground in liquid nitrogen, followed by extraction with a hot cetyltrimethylammonium bromide (CTAB) buffer, phase separation using chloroform, and RNA precipitation with lithium chloride. Ethanol washes ensure purity before resuspension in RNases free water. Key reagents such as CTAB, polyvinylpyrrolidone, Ethylenediaminetetraacetic acid (EDTA), Spermidine, and β-mercaptoethanol enhance RNA yield and quality. Completed within approximately 36 h, the protocol yields between 120 and 2120 ng/μL, with leaf tissues generally providing the highest amounts. This optimized method delivers high-integrity RNA, making it well-suited for diverse downstream molecular applications.