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Add Trizol to cells and lysate the cells by mixing the mixture thoroughly; stand the mixture for 5 minutes and then isolate RNA immediately at room temperature or store the lysates at -80C until further analysis. For stored samples, thaw the lysates completely and add 200 ul of chloroform/ml Trizol. Mix the mixture completely by inversion repeatedly for 30s and stand for 3 minutes at room temperature, followed by centrifugation at 12,000 x g for 15 min.
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