user's manuals about RNA or or

NucleoZOL

2.2 Product specificationsTable 1: Product specifications at a glance Technology Sample material (per mL NucleoZOL)7

One-phase extraction< 1 x 10 cultured cells, bacteria, and yeast,< 100 mg human/animal/plant tissue,< 0.4 mL (viral) fluids Small RNA (10–200 nt), large RNA (> 200 nt) Liver: 6–8 μg/mg tissue Kidney, spleen: 3–4 μg/mg tissue Muscle, brain, lung: 0.5–1.5 μg/mg Cultured cells: 4–10 μg/106 cells

Fragment size Typical yield (total RNA)

Typical yield (large RNA)

Liver: 5–7 μg/mg tissue Kidney, spleen: 3–4 μg/mg tissue Muscle, brain, lung: 0.5–1.5 μg/mg Cultured cells: 3–8 μg/106 cells

A260/280 (total RNA) Typical RIN (RNA integrity number) Elution volume

1.8–2.1> 9 flexible

MACHEREY-NAGEL– 02/2016, Rev. 02

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user's manuals about RNA or or

2.3 Handling, preparation, and storage of starting materials

Sample harvest and RNase inhibition

RNA is not protected against digestion until the sample material is flash frozen or disrupted in the presence of RNase inhibiting or denaturing agents. Sample harvest methods:

Use freshly harvested sample for immediate lysis and RNA purification.Samples can be stored in NucleoZOL after disruption at -20 °C to -70 °C for up to one year, at 4 °C for up to 24 hours, or up to several hours at room temperature. Frozen samples in NucleoZOL should be thawed slowly before starting with the isolation of RNA.

Flash freeze sample in liquid N2 immediately upon harvest and store at -70 °C. Frozen samples are stable up to 6 months. Mortar and pestle can be used to pulverize the sample in a frozen state. Make sure that the sample does not thaw prior to contact with the reagent.

Samples can be submerged and stored in RNA stabilizing reagents such as RNAlater®. Remove excess RNAlater® solution from the tissue before processing the sample.

2.4 RNA reconstitution

The precipitated RNA can be dissolved in variable volumes of RNase-free water (see ordering information, section 6.3) to approach an RNA concentration of approximately 1–2 μg/μL for the large RNA fraction and approximately 0.1 μg/μL for the small RNA fraction.