Qiagen, 954500, Buffer RPP (8mL)

The EZ2 RNA/miRNA Tissue/Cells Kit makes it easy and safe to purify total RNA – including miRNA and other small RNA – from cultured cells and various human or animal tissues. The process requires no use of phenols, and once the lysis step is completed, all remaining steps are automated with any EZ2 Connect instrument.
The EZ2 RNA/miRNA Tissue/Cells protocol isolates a similar amount of total RNA, including small RNA, to manual column-based protocols. This is true even for difficult tissue types such as fibrous tissue (e.g., muscle) or fatty tissue (e.g., brain) (see Comparable miRNA recovery). Genomic DNA removal is likewise comparable to manual column-based purification (see Efficient mRNA recovery and gDNA removal). Digital PCR analysis shows higher RNA concentration in samples processed with the EZ2 RNA/miRNA Tissue/Cells Kit (see Higher RNA concentration).
The EZ2 RNA/miRNA Tissue/Cells protocol uses manual lysis in Buffer RLT protect the RNA molecules. Proteinase K digestion ensures complete lysis of even hard-to-lyse tissues. Bind, wash, and elute steps are automated on an EZ2 Connect instrument, which uses magnetic bead technology. After first binding, RNA is treated with DNase to digest contaminating genomic DNA, and then rebound to the magnetic beads. Wash steps then remove contaminants that could interfere with enzymatic reactions that follow.
The cell pellet or cell culture is first homogenized, or the tissue sample is disrupted and then homogenized. RNase-free water and Proteinase K are then added, and the mixture is incubated for 10 minutes at room temperature (see EZ2 RNA/miRNA Tissue/Cells workflow). After incubation, samples and prefilled reagent cartridges are loaded into the EZ2 Connect, and application run is started. Once the run is finished, purified RNA or miRNA will be ready for downstream applications (see EZ2 RNA/miRNA Tissue/Cells workflow).