Abstract: 【OBJECTIVE】To find the best methods for isolating high quality total RNA from different tissues of Tobacco.【METHOD】 Three methods, which are the kit, the Trizol regent and the CTAB for RNA isolation, were used to compare total RNA of 6 tissues .【RESULTS】The results show that there are different effects of 3 methods for isolating RNA form different tissues of Tobacco. The kit and the Trizol regent for RNA isolation aren’t suitable for isolating total RNA from Tobacco seed, while the CTAB method can be isolated total RNA from every Tobacco tissues. Moreover, the RNA can be used for the RT-PCR and the other subsequent experiment. 【CONCLUSION】The kit is simple and quick method for isolation RNA from Tobacco, compared with the Trizol regent and the CTAB method.