user's manuals about RNA or or

2

Precipitate contaminants

Add 400 μL RNase-free water per 1 mL NucleoZOL to the lysate. Shake the sample vigorously for 15 s. Incubate at room temperature (18–25 °C) for 5–15 min.

For samples containing 100 mg tissue/mL NucleoZOL, 15 min incubation at room temperature is recommended.

Centrifuge samples for 15 min at 12,000 x g. Centrifugation can be performed at 4–28 °C.

A semi-solid pellet containing DNA, proteins, and polysaccharides forms at the bottom of the tube. The RNA is still solubilized in the supernatant. 3

Precipitate large RNA

Pipette 1 mL of the supernatant to a new tube. Leave a layer of the supernatant on top of the precipitate. Add 400 μL 75 % ethanol to 1 mL supernatant for precipitation of the RNA.

Incubate samples at room temperature for 10 min.

Centrifuge the samples for 8 min at 12,000 x g. A white pellet containing the RNA will be formed at the bottom of the tube. Transfer the supernatant containing the small RNA to a new tube and store it at 4 °C or at -20 °C.

The small RNA containing supernatant can be stored at -20 °C for one year.4

Precipitate small RNA

Add 0.8 volumes of isopropanol to the supernatant (~ 1.4 mL) obtained after precipitation of RNA (step 3) Incubate the samples for 30 min at 4 °CCentrifuge the sample for 15 min at 12,000 x g. Centrifugation can be performed at 4–28 °C.

Precipitated RNA will form a white pellet at the bottom of the tube.