A Simple and efficient protocol for isolation of RNA from different tissues of chickpea and pea

Abstract

A simple and efficient protocol is developed for isolation of high quality RNA from roots and leaves of chickpea and pea. The procedure is based on use of SDS, sodium acetate and EDTA in an extraction buffer in order to eliminate polysaccharides and prevent oxidation of phenolic compounds. The current method is modification of a method described for RNA isolation from pea leaves only, and yields excess amount of high-quality RNA suitable for cDNA based gene expression analysis. The protocol requires only three disposable micro centrifuge tubes during extraction, single phenol extraction step and a single precipitation step to yield high-quality RNA. RNA extracted with this method was free from protein and phenolic contaminants as evident from gel electrophoresis analysis. This method is applicable not only for leaves but also for roots and shoots and equally applicable to both chickpea and pea. cDNA is prepared and PCR amplification have been done with universal ubiqutin primer to check the integrity of RNA and absence of inhibitory compounds in RNA samples, which proves the suitability of samples towards qRTPCR.