NucleoSpin FFPE RNA is designed for the isolation and purification of total RNA from formalin-fixed, paraffin-embedded (FFPE) samples. In the first stage, paraffin is removed from the sample, utilizing the odor-free Paraffin Dissolver. Protocols are available in the user manual for users preferring xylene deparaffinization. Deparaffinization using xylene shows the same efficiency as deparaffinization using Paraffin Dissolver. After deparaffinization, the sample is lysed, releasing the nucleic acids. Incubation of the RNA with D-Link Buffer effectively removes crosslinking from RNA and reverses formaldehyde modifications. The decrosslinking step takes only 15 minutes and RNA can be purified by binding to the NucleoSpin FFPE column.
NucleoSpin FFPE RNA kits allow elution of RNA in as little as 5 µl, resulting in higher concentrations compared to other manufacturer’s kits. NucleoSpin FFPE RNA columns are designed for increased sensitivity. A silica membrane of small diameter is embedded in a funnel-shaped ring (see column cross-section), allowing an elution volume of only 5–30 µl.
Isolated RNA has a high concentration, formaldehyde crosslinks are removed efficiently, and RNA integrity is well preserved during the complete process (deparaffinization, decrosslinking, purification). Thus, RNA shows good performance in typical downstream applications like RT-PCR.