International Journal of Molecular and Cellular Medicine (IJMCM)

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The reliability of gene expression profiling based technologies and methods to find transcriptional differences representative for the original samples is influenced by the quality of the extracted RNA. Hence, RNA extraction is the first step to investigate the gene expression and its function. Consequently, the quality of extracted RNA is really significant. Correspondingly, this research was accomplished in order to optimize the RNA extraction methods and compare the tissue amounts or tissues quality. Relatively, the cancerous tissues of human's stomach in fresh and frozen conditions and also the mouse's fresh tissue were studied. Some factors like the amount of samples, efficacy differences of diverse extraction's buffers (TriPure, Trizol) and also the efficacy of b-mercaptoethanol were compared and investigated. The results indicated that the less amount (1-2 mg) compared to other amounts (2-5 mg, 5-15 mg) yielded the best quality and the RNA's bands (5S, 18S, 28S) were observed perfectly. Relatively, comparing and measuring some kinds of buffers (Trizol, TriPure) indicated no difference in RNA extraction quality. The last investigated factor was the effect of b- mercaptoethanol which was used along with TriPure in order to remove the RNAse. Conclusively, no effective impression was observed.

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