TRIzol Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it.

Protocol tips
- For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it.

Publication protocol

For transcriptome analysis, RNA from DAOY cells was extracted by using Trizol (Life Technologies). Briefly, cells were washed with phosphate buffer saline (PBS) and lysed in 1 ml Trizol. Chloroform (0.2 ml) was added, and the samples were mixed intensively and incubated for 5 min at room temperature. The upper aqueous phase was transferred to a new tube after centrifugation (12 000 g, 15 min, 4 °C) and mixed with 0.5 ml of isopropanol. After incubation at 4 °C for 10 min, the precipitated RNA was pelleted by centrifugation (12 000 g/15 min/4 °C) and washed with 75 % ethanol. The RNA pellet was dissolved in 20 µl of RNase-free water. The RNA was stored at −80 °C until further use.

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