For purification of total RNA from formalin-fixed paraffin embedded (FFPE) tissue sections for in vitro diagnostic use

Features

    Isolates RNA from FFPE tissue sections and includes Deparaffinization Solution
    Efficiently removes formalin crosslinking
    Releases RNA without compromising RNA integrity
    FFPE RNA purification system for use with other IVD products

Product Details
The RNeasy DSP FFPE Kit is specially designed for purifying total RNA from FFPE tissue sections. The Deparaffinization Solution removes paraffin from the tissue sample. Special lysis and incubation conditions reverse formaldehyde modifications of RNA. In addition, the lysis buffer ensures efficient release of total RNA from tissue sections, while avoiding further RNA degradation. The kit also uses DNase and DNase Booster Buffer to optimally remove genomic DNA contamination. RNeasy MinElute columns, also included within the kit, enable total RNA elution in as little as 14 μl.

Principle
Fixing tissues with formalin leads to RNA-RNA and RNA-protein crosslinking which impairs RNA performance in enzymatic assays. Fixation and embedding conditions can also result in heavily fragmented nucleic acids in FFPE samples. The RNeasy DSP FFPE Kit uses special lysis and incubation conditions to reverse formaldehyde modifications of RNA. The lysis buffer efficiently releases RNA from FFPE tissue sections, preventing further RNA degradation. These optimized conditions allow purification of all RNA, leading to greater yields of RNA from FFPE samples.

Procedure
The RNeasy DSP FFPE Kit uses spin-column based technology to purify RNA from FFPE tissue sections. Paraffin is removed from tissue sections using Deparaffinization Solution. Proteinase K is then added to the sample for lysis. DNase Booster Buffer and DNase I are added for removal of genomic DNA. Ethanol is added to the sample and transferred to the RNeasy MinElute column. Subsequent washing and centrifugation steps are applied before total RNA is eluted in 14–32 µl RNase-free water.

Applications
The RNeasy DSP FFPE Kit isolates total RNA from FFPE samples, providing highly pure RNA fragments for use in a wide range of downstream applications. Please note that due to the fixation treatment, RNA purified from FFPE tissue sections can be heavily fragmented, which might impair the usability in downstream applications requiring full-length RNA.