Optimization of RNA extraction protocol for trees bark tissue of coniferous family

RNA extraction from hard wood tissues, especially in coniferous family trees, is very hard because they have high concentrations of phenolic compounds, polysaccharides, secondary metabolites and lignin. Moreover, up to now, no commercial kit is made for the mentioned purpose. In this study, the problems were resolved and a rapid, simple, and economic protocol is presented for RNA extraction from bark tissue of Cedrus deodara and Abies grandis, which belong to conifer family. Ratios of optical density of 260/280 nm and 260/230 nm were respectively obtained as 1.88 and 1.1 for Cedrus deodara and 1.8 and 0.9 for Abies grandis. Modifications for optimization of RNA extraction were preheating treatment of extraction buffer, and decreasing centrifuge rounds to eliminate DNA contaminations and DEPC. Synthesis of cDNA was performed using extracted RNA. RT-PCR successfully produced expected bands. Therefore, it seems that the presented protocol is suitable for RNA extraction from woody tissues.