TRI Reagent™ Solution

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Carefully watch how much TRI reagent should be added to your samples.

Upstream tips
- Carefully watch how much TRI reagent should be added to your samples.

Publication protocol

After 7, 14, or 21 days of culturing, total RNA was extracted from the samples using TriReagent according to the manufacturer's instructions. In brief, ice-cold Tri Reagent (1 mL) was added to the samples, and they were incubated for 5 min at room temperature. Then, 1-bromo-3-chloropropane (BCP; 0.1 mL) was added, and they were incubated for a further 15 min at room temperature. The samples were centrifuged at 12,000× g for 15 min at 4°C, and the upper aqueous phase that contained the RNA was collected. The RNA was precipitated using isopropanol and centrifuged at 12,000× g for 10 min at 4°C. Following precipitation, 1 mL of 75% ethanol was added into the RNA pellet, followed by centrifugation at 7500× g for 5 min, and the resulting pellet was air dried. The RNA pellet was dissolved in DEPC water. The concentration and purity of each sample were assessed by absorbance at 260 nm and by the 260/280 nm ratio, respectively.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for TRI Reagent™ Solution below.

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