Description
The QIAsymphony RNA Kit enables automated purification of total RNA, or total RNA including miRNA and other small RNAs, from 1–96 samples on the QIAsymphony SP. Total RNA can be purified from animal cells, tissues, or FFPE tissues, using very small amounts of starting material (≥100 cells) up to 1 x 10 7 cells or 50 mg of tissue. Optimized protocols provide high yields of pure total RNA, or total RNA including miRNA, and flexible elution volumes allow RNA concentration to be easily optimized for each downstream application. Prefilled reagent cartridges reduce setup time and minimize manual handling. Bar code reading enables full tracking of reagents.
Automated purification of RNA on the QIAsymphony SP is highly reproducible and yields of high-quality RNA from cultured cells, tissues, and FFPE tissues are comparable to those obtained using a manual purification procedure (see figure "Reproducible purification of RNA"). The QIAsymphony protocols for total RNA purification primarily enrich for RNA molecules longer than 200 nucleotides, including mRNA (see figure "Highly pure, intact RNA"). The QIAsymphony miRNA protocol also purifies total RNA but does not exclude RNAs <200 nucleotides such as miRNAs and other small RNAs (see figure "Automated purification comparable to manual procedure").
The QIAsymphony RNA Kit combines the speed and efficiency of silica-based technology with the convenient handling of magnetic particles (see figure "Efficient handling of magnetic particles") for purification of total RNA >200 nucleotides, or total RNA including miRNA and other small RNAs after proteinase K treatment. Starting material includes animal cells and tissues. With some additional components, formalin-fixed paraffin-embedded (FFPE) tissues can also be processed with the kit. Samples are lysed and RNA is purified from lysates in one step through its binding to the silica surface of the particles in the presence of a chaotropic salt. The particles are separated from the lysates using a magnet and DNA is removed by treatment with RNase-free DNase. The magnetic particles are efficiently washed, and RNA is eluted in RNase-free water.
Innovative, ready-to-run reagent cartridges are prefilled with magnetic particles and all reagents required for the purification procedure, including accessory enzymes. The cartridges are automatically opened by the QIAsymphony SP helping to minimize the risk of environmental contamination (see figure "Prefilled, sealed reagent cartridges"). Worktable setup is rapid, which saves your valuable time. Simply place up to 2 reagent cartridges in the consumables drawer. The reagent cartridges can be either from the same kit or from different kits, enabling different purification procedures to be performed within the same run of 96 samples. Reagent volumes only for the selected number of samples are used, giving you complete cost control. The purification procedure comprises 4 steps: lyse, bind, wash, and elute. A number of protocols is available for the QIAsymphony SP for purifying RNA, depending on the starting material and the required RNA product (see table). Protocols for purification of RNA with the QIAsymphony SP instrument Protocol Running with Sample material (Pretreated sample lysates) Processed volume (μl) RNA CT 400 QIAsymphony RNA Kit (2 x 96) ‡ ≤3 x 10 6 cultured cells ≤20 mg easy-to-lyse tissue 400 μl RNA CT 800 QIAsymphony RNA Kit (2 x 48) ‡ 3 x 10 6 –1 x 10 7 cultured cells ≤50 mg easy-to-lyse tissue 800 μl RNA FT 400* QIAsymphony RNA Kit (2 x 96) ‡ ≤20 mg fibrous tissue 400 μl miRNA CT 400* QIAsymphony RNA Kit (2 x 96) ‡ ≤3 x 10 6 cultured cells ≤20 mg tissue 400 μl RNA FFPE 130 † QIAsymphony RNA Kit (2 x 72) ‡ 1–2 x FFPE sections up to 10 μm 1 x FFPE section 20 μm 130 μl * Proteinase K required; can be ordered separately. † Additional components required; can be ordered separately. ‡ Values in parentheses indicate maximum number of samples that can be processed with kit/protocol combination.
