Background Plant tissues must be preserved in their collection state, especially for genome-wide expression profile studies. Lyophilization is a feasible, affordable tool when fresh tissues cannot be shipped at ultralow temperatures from their origin to the place of analysis. In this study, the total RNA quality of dormant grapevine buds (Vitis vinifera L. cv. Flame Seedless ) of freeze-dried samples stored at room temperature conditions was evaluated and compared to that of cryopreserved (-80°C) grapevine buds. Results Good yield and quality of RNA were obtained from freeze-dried dormant buds stored at room temperature for 0, 3 and 6 weeks after they were lyophilized. Further experiments confirmed that the extracted total RNA could be used for actin and β-tubulin PCR gene amplification. Conclusion High-quality RNA that is useful for downstream applications was obtained from freeze-dried dormant grapevine bud tissue, similarly to the RNA obtained from cryopreserved dormant grapevine buds.