Isolation of RNA from Grains of Medicinal Rice “Njavara” using Improved TRIzol Method

Isolation of high-quality RNA from seeds is very critical for seed specific gene analysis. An investigation and comparison on the use of CTAB, SDS and TRIzol extraction procedures to extract high-quality RNA from grains of medical njavara rice were carried out in the present study. These protocols either failed to yield RNA or resulted in reduced yield with poor quality of RNA from Njavara rice grains. The starch and secondary metabolites present in njavara rice seeds hindered the isolation and the resuspension of precipitated RNA or contaminated the RNA pellets by co-precipitation. Hence, we modified the TRIzol RNA extraction protocol by addition of 0.5% N-lauryl sarcosine, 2% β-Mercaptoethanol and 1% PVP. Highly pure (A260/A280 ratio ranged from 1.9 to 2.0) and intact RNA with a higher yield (up to 500µg/ml of RNA) could be obtained using modified TRIzol RNA extraction protocol. RNA obtained was testified and efficiently utilized for the cDNA preparation and amplification of Ubiquitin gene.