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42
RNeasy Mini Handbook 10/2019
The following guide may help you to determine the amount of RNAprotect Tissue Reagent
required for RNA stabilization:
A cube of rat kidney with a 5 mm edge length ([5 mm]
3
= 125 mm
3
= 125 µl) weighs
150–175 mg and requires at least 1.5–1.75 ml of the reagent.
A 3 mm cube ([3 mm]
3
= 27 mm
3
= 27 µl) of most animal tissues weighs 30–35 mg and
requires at least 300–350 µl of the reagent.
Although weighing tissues is generally more accurate, RNA in unstabilized tissues will degrade
during weighing. In some cases, however, it may be more convenient to quickly estimate the
weight of tissue pieces. Average weights of various entire adult mouse organs and the
corresponding amounts of RNAprotect Tissue Reagent to use are given in Table 7.
Table 7. Tissue Weights and Amounts of RNAprotect Tissue Reagent
Important points before starting
If using the RNeasy Protect Mini Kit for the first time, read “Important Notes” (page 19).
RNAprotect Tissue Reagent may form a precipitate during storage below room
temperature. Before using the reagent, redissolve the precipitate by heating to 37°C with
agitation.
Only fresh, unfrozen tissues can be stabilized using RNAprotect Tissue Reagent.
Previously frozen tissues thaw too slowly in the reagent, preventing the reagent from
diffusing into the tissues quickly enough to prevent RNA degradation.
Mouse organ Weight (mg)
Amount of RNAprotect
Tissue
Reagent (ml)
Kidney 180–250 ≤2.5
Spleen 100–160 ≤1.6
Lung 190–210 ≤2.1
Heart 100–170 ≤1.7
Liver 1000–1800 ≤18
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