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RNeasy Mini Handbook 10/2019
45
Protocol: Purification of Total RNA from Animal
Tissues
This protocol requires the RNeasy Mini Kit or RNeasy Protect Mini Kit.
Determining the correct amount of starting material
It is essential to use the correct amount of starting material in order to obtain optimal RNA
yield and purity. A maximum amount of 30 mg fresh or frozen tissue or 15–20 mg RNAprotect
stabilized tissue (which is partially dehydrated) can generally be processed. For most tissues,
the RNA binding capacity of the RNeasy spin column and the lysing capacity of Buffer RLT
will not be exceeded by these amounts. Average RNA yields from various tissues are given in
Table 2 (page 21).
Some tissues, such as spleen, parts of brain, lung, and thymus are more difficult to lyse or tend
to form precipitates during RNA purification. The volume of Buffer RLT may need to be
increased to facilitate complete homogenization and to avoid significantly reduced RNA
yields, DNA contamination, or clogging of the RNeasy spin column. See the procedure below
for details.
RNA yields from fibrous tissues, such as skeletal muscle, heart, and skin, may be low due to
the abundance of contractile proteins, connective tissue, and collagen. For maximum RNA
yields from these tissues, we recommend using the RNeasy Fibrous Tissue Mini Kit instead.
See page 91 for ordering information.
Greater RNA yields from fatty tissues, such as brain and adipose tissue, can be achieved using
the RNeasy Lipid Tissue Mini Kit, which uses QIAzol Lysis Reagent for optimal tissue lysis. See
page 91 for ordering information.
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