UM_TotalRNA_NucleoZOL(14)

user's manuals about RNA or or

5

Wash RNA

Use 400–600 μL 75 % ethanol when precipitating in 1.5 mL tubes. For larger tubes, add 500 μL 75 % ethanol per 1 mL supernatant

Centrifuge the pellets for 1–3 min at 4,000–8,000 x g. Remove ethanol from the pellet by pipetting. Repeat the ethanol washing step. Do not dry the RNA pellet between wash steps.

Drying the RNA pellet may lead to a decrease in solubility.6

Reconstitute RNA

Dissolve the RNA pellet in RNase-free water to obtain an RNA concentration of 1–2 μg/μL. Vortex the sample 2–5 min at room temperature for efficient solubilization.

For accurate determination of RNA concentration by OD measurement with a cuvette, dilute RNA in RNase-free water with a slightly alkaline pH, 1 mM NaOH or buffer with a pH > 8 (e.g., Elution Buffer AE, see ordering information, section 6.3). Distilled water typically has a pH < 7.