An efficient and rapid method for the isolation of RNA from different recalcitrant tissues of mango (Mangifera indica L.).

Abstract

The isolation of high-quality RNA from different tissues of mango (Mangifera indica L.) is relatively challenging due to the presence of interfering substances such as polysaccharides, polyphenols, and proteins. All these compounds render available isolation protocols useless by reducing the quality (purity and integrity) and quantity of the RNA that can be recovered. Several tissue-specific protocols for the isolation of RNA have been developed specifically for mango, however, they are cumbersome, expensive and time-consuming. To overcome these drawbacks, we have developed a comprehensive (CTAB-free, guanidine-free, and LiCl-free) RNA isolation protocol using SDS (sodium dodecyl sulphate) plus phenol which works well for most mango tissues such as leaves, flowers, and fruit, at different stages of development or ripening, as well as fruit peel and seed kernels.This rapid protocol allowed us to process large numbers of samples (12 – 15) simultaneously in a single day. Using this method, we obtained good quantity RNA (16 – 80 µg g–1 tissue) from various mango tissues at different stages of development. RNA isolated by this method was pure and amenable to various downstream molecular applications such as RT-PCR and the construction of a cDNA library.