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Isolation of high-quality RNA from different strawberry tissues is often affected by the presence of high levels of contamination by polysaccharides and phenolic compounds. By the methods of improved CTAB, SDS, and guanidinium thiocyanate, total RNA was isolated from leaves of strawberry. The result indicated that total RNA is of high quality and undegraded by using CTAB. However, it was difficult to isolate total RNA from leaves of strawberry by using guanidinium thiocyanate, and total RNA could be extracted by using SDS with low concentration, impurity and degradation in some degree. After comparing, an improved 3% CTAB 3 was used to isolate RNA from tissues of strawberry, and the isolated RNA was good enough for further applications. After RT-PCR, the 699 bp sequence of glucose-6-phosphate dehydrogenase (G6PDH) gene was obtained to identify the quality of total RNA from strawberry. In this experiment, improved 3% CTAB 3 method could be regarded as a simple, rapid, economic and convenient method for the RNA isolated from strawberry.