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RNeasy Fibrous Tissue Mini Handbook 01/2018
Introduction
The RNeasy Fibrous Tissue Mini Kit is designed for optimal lysis of fiber-rich tissues and
purification of high-quality total RNA >200 nt.
Principle and procedure
Total RNA purification from fibrous tissues, such as skeletal muscle, heart and aorta tissue can
be difficult due to the abundance of contractile proteins, connective tissue and collagen. The
RNeasy Fibrous Tissue Mini Kit is supplied with proteinase K, which removes these proteins.
Tissue samples are first lysed in Buffer RLT and then diluted before being treated with
proteinase K. Debris is pelleted by centrifugation, and the supernatant is removed. The
supernatant is mixed with ethanol and then centrifuged through an RNeasy Mini spin column,
where RNA binds to the silica membrane. Traces of DNA that may co-purify with the RNA are
removed by DNase treatment on the silica membrane. DNase and any contaminants are
efficiently washed away, and high-quality total RNA is eluted in RNase-free water. The RNeasy
Fibrous Tissue Mini Kit purifies RNA from up to 30 mg tissue (Figure 1, next page).
With the RNeasy Fibrous Tissue Mini Kit, all RNA molecules longer than 200 nucleotides are
purified. The procedure provides enrichment for mRNA since most RNAs <200 nucleotides
(such as 5.8S rRNA, 5S rRNA, and tRNAs, which together comprise 15–20% of total RNA)
are selectively excluded. For purification of small RNA, including microRNA, from tissues
and cells, we recommend using miRNeasy Kits.
