RNeasy FFPE Kit

Total RNA was purified from a 10 µm rat lung FFPE sample using the RNeasy FFPE Kit or alternative kits from Supplier E_III, Supplier N, Supplier A_IV, and Supplier M. Real-time RT-PCR assays for PGK1 (phosphoglycerate kinase 1) were performed on the Rotor-Gene Q cycler with (+RT) or without (-RT) the reverse transcription step. The high C_T value in the -RT sample from the RNeasy FFPE Kit indicates that purified RNA is virtually free of genomic DNA. Therefore the C_T value in the +RT sample from the RNeasy FFPE Kit is a reliable measure of RNA expression.|RNA was purified from 10 µm FFPE sections of the indicated rat tissues using either the RNeasy FFPE Kit or alternative kits from Supplier N, Supplier A_IV, and Supplier M. RNA yields were determined by measuring absorbance at 260 nm.|After paraffin is removed from samples, the optimized lysis buffer allows sample lysis with proteinase K digestion in only 15 minutes without sacrificing RNA yields. After lysis, samples are incubated at 80ºC for 15 minutes to reverse formalin crosslinking. Genomic DNA, including small DNA fragments found in FFPE samples, is then effectively removed using DNase and DNase Booster Buffer. Finally, concentrated RNA is purified using RNeasy MinElute spin columns. Since RNA is eluted in a volume of just 14–30 µl, smaller reaction volumes are possible in downstream applications.|Total RNA was purified from rat kidney using the RNeasy FFPE Kit. Real-time RT-PCR assays for PGK1 (phosphoglycerate kinase 1) were performed on the Rotor-Gene Q cycler with (+RT) or without (-RT) reverse transcriptase. The -RT curves demonstrated that RNA purified using the RNeasy FFPE Kit was virtually free of genomic DNA.|RNA purified from 6-month-old FFPE rat liver using the RNeasy FFPE Kit or alternative kits from Supplier A_IV, Supplier N, or Supplier M, was analyzed on the Agilent 2100 Bioanalyzer. The peaks indicate that more intact RNA was purified using the RNeasy FFPE Kit than when using alternative kits.|