Principle

The NucleoSpin® totalRNA FFPE kit provides a convenient, efficient, and fast method to isolate large and small RNA from standard formalin-fixed, paraffin-embedded (FFPE) tissues. In the first step paraffin is removed from  sample with the odorless and non-toxic blue Paraffin Dissolver (patent pending). The blue color visualizes the Paraffin Dissolver for an easy and convenient handling. Decrosslinking and tissue lysis is done in one step providing reliable total RNA yields.
Optimal binding conditions for even small RNA (e.g., miRNA) are adjusted and the lysate is applied to the NucleoSpin®RNA Column. RNA is bound to the silica membrane. Residual DNA is removed by the convenient on-column rDNase digestion. The washing steps remove salts, metabolites, and macromolecular cellular components. Total RNA is finally eluted in 30-50 µL.

Features
 

Application data

Excellent RT-PCR performance and most efficient gDNA removal with NucleoSpin® totalRNA FFPE
Large (e.g., mRNA) and small (e.g., miRNA) RNA was isolated from 4 x 10 μm FFPE sections of mouse brain tissue with NucleoSpin® totalRNA FFPE compared to three other competitor kits (Q, A, P). (A) Quantification of mRNA* and miRNA** was done by qRT-PCR. Low CT values indicates high RNA yields. (B) Residual DNA was assayed by amplifying a 191 bp fragment of the mGAPDH gene. The higher the CT values the lower the residual DNA.

*Target length 230 bases of the β2-microglobulin gene
**Applied Biosystems, TagMan® MicroRNA RT Kit, hsa-miR-16 MicroRNA Assay