QIAsymphony PAXgene Blood RNA Kit

The fully automated PAXgene Blood RNA procedure provides RNA yields comparable to the manual PAXgene Blood RNA procedure. Typical yields of RNA isolated from 2.5 ml healthy human whole blood (4.6 x 10⁶ - 1.1 x 10⁷ leukocytes/ml) are ≥3 μg for >95% of the samples processed with all PAXgene Blood RNA Kits. The automated PAXgene Blood RNA procedure provides high-quality RNA from different donor samples with enrichment of small RNA species (see figure "High-quality RNA from different donors"). Since yields are highly donor-dependent, individual yields may vary. The integrated DNase treatment ensures that genomic DNA contamination is minimized. Purified RNA is ready to use in downstream applications, such as quantitative, real-time RT-PCR.

The QIAsymphony PAXgene Blood RNA Kit provides fully automated purification of intracellular RNA on the QIAsymphony SP. Used with PAXgene Blood RNA Tubes (available from BD and BD authorized distributors, cat. no. 762165) for sample collection and stabilization, the system provides a complete preanalytical solution from sample collection and stabilization through to purification of high-quality RNA using proven magnetic-particle technology. Prefilled cartridges reduce manual handling to a minimum, and bar code labeling enables full reagent tracking.

The QIAsymphony PAXgene Blood RNA procedure begins with collection of blood samples (2.5 ml) in PAXgene Blood RNA Tubes. Samples can be stored or transported at room temperature. The purification procedure starts with a centrifugation step to pellet the contents of each PAXgene Blood RNA Tube. After resuspension of the pellet, the tubes are transferred to the QIAsymphony SP. Loading of the reagent cartridge and plasticware is guided and checked by dedicated software. The QIAsymphony SP then carries out the fully automated RNA purification procedure. An integrated DNase treatment ensures that genomic DNA contamination is minimized (see figure "QIAsymphony PAXgene Blood RNA procedure"). The robust and standardized procedure together with on-board measures of the QIAsymphony SP is designed to minimize the risk of sample-to-sample cross-contamination (see figure "On-board measures against cross-contamination"). After purification, a final heat treatment of the eluate enhances performance in downstream applications. Total processing time is about 2.5 hours for 24 samples, which includes a maximum of 25–30 minutes of hands-on time.

RNA purified using the QIAsymphony PAXgene Blood RNA Kit and the QIAsymphony SP is highly pure and ready for use in a wide range of sensitive downstream applications, including:

• RT-PCR and quantitative, real-time RT-PCR
• Expression-array and expression-chip analysis cDNA synthesis
• RNase and S1 nuclease protection
• Northern-, dot-, and slot-blot analysis