QIAsymphony PAXgene Blood RNA Kit

For automated purification of intracellular RNA (including miRNAs) from stabilized blood

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QIAsymphony PAXgene Blood RNA Kit (96)

Cat. No. / ID: 762635

For 96 preps: 2 Reagent Cartridges, Enzyme Racks, accessories, and RNase-free buffers

For Research Use Only. Not for use in diagnostics procedures. No claim or representation is intended to provide information for the diagnosis, prevention, or treatment of a disease.

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Features

Improved RNA quality and yield
Effective copurification of miRNA
Prefilled reagent cartridges for safety and ease of use
High reproducibility through standardized processing
Bar codes enable tracking of samples and reagents

Product Details

The QIAsymphony PAXgene Blood RNA Kit provides fully automated purification of cellular RNA on the QIAsymphony SP. The new, improved kit provides higher-quality RNA, including miRNA. 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.

Performance

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.

See figures

High-quality RNA from different donors.

Principle

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.

Procedure

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 ""). 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 ""). 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.

See figures

QIAsymphony PAXgene Blood RNA procedure.

On-board measures against cross-contamination.

Applications

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

Supporting data and figures

High-quality RNA from different donors.

Blood was collected from 12 donors into PAXgene Blood RNA Tubes. RNA was purified using the QIAsymphony PAXgene Blood RNA Kit on the QIAsymphony SP. RNA was analyzed with an Agilent 2100 Bioanalyzer. Clear enrichment of small RNA species is shown in the gel-like image by the corresponding peaks in the donor RNA next to the marker band.

Specifications

Features	Specifications
Applications	Quantitative, real-time RT-PCR, expression array and chip analysis, next-generation sequencing, cDNA synthesis, RNase and S1 nuclease protection, northern, dot, and slot blot analysis, primer extension
Time per run	72 samples in 6 h 15 min
Processing	Automated on the QIAsymphony
Format	Magnetic beads
Sample amount	2.5 ml
Yield	8–16 µg
Main sample type	Whole blood
Elution volume	80 µl or 120 µl or 200 µl
Technology	Magnetic particles technology

Resources

For collection, transport, and storage of whole blood and stabilization and purification of intracellular RNA

For collection of stable blood samples and purification of high-quality RNA for demanding downstream applications

For use with software version 5.0 or higher

For purification of total RNA, including miRNA, from human whole blood stabilized in PAXgene Blood RNA Tubes using the QIAsymphony SP and QIAsymphony PAXgene Blood RNA Kit.

Typical total RNA yields from PAXgene Blood RNA Tubes processed with the QIAsymphony PAXgene Blood RNA Kit

Voss et al., AMP 2010

FAQ

Room temperature storage of the filled PAXgene Blood RNA Tubes for up to three days has no effect on the RNA profile. For longer storage they should be stored at –20°C or –70°C.

FAQ ID -2984

RNA yields isolated from 2.5 ml healthy, human, whole blood (4.8 x 10⁶ –1.1 x 10⁷ leukocytes/ml) are specified to ≥3 µg for >95% of the samples processed (see Technical Note for the QIAsymphony PAXgene Blood RNA System ). Since yields are highly donor-dependent, individual yields may vary. Mean yields in the range of 8-10µg per tube or higher.

FAQ ID - 3475

Stability data are not yet available. A study is ongoing.

FAQ ID -2983

Denaturation of the eluate is essential for maximum efficiency in downstream applications, such as RT-PCR, other amplification reactions, or cDNA synthesis. It is not necessary to denature samples more than once; samples remain denatured after freezing and thawing.

FAQ ID -2987

We recommend using Elution Microtubes CL, racked (provided) for elution. Alternatively, 2 ml Sarstedt tubes can be used. Other elution formats will be supported in the future, so please refer to the current labware list.

FAQ ID -2986

The product is not for the isolation of genomic DNA or viral nucleic acids from human whole blood.

FAQ ID - 3476

Due to limited plastic consumables on board the instrument, the number of batches for the QIAsymphony PAXgene Blood RNA protocol is limited to three at one time. Therefore up to 72 samples can be processed in a single run.

FAQ ID -2498

Yes, samples with the QIAsymphony PAXgene Blood RNA Protocol have to be incubated at least 2 hours at room temperature (18– 22°C). Incubation of the PAXgene Blood RNA Tube overnight may increase yields slightly in some cases. Room temperature storage for up to 3 days may have no effect on the RNA profile. During the preparation of the RNA the manual pretreatment consists of a centrifugation and a pellet resuspension step (see the QIAsymphony PAXgene Blood RNA Kit Handbook).

FAQ ID -2982

No. The QIAsymphony PAXgene Blood RNA System is ’For Research Use Only’. Not for use in diagnostic procedures. No claim or representation is intended to provide information for the diagnosis, prevention, or treatment of a disease. The performance characteristics of this product have not been fully established.

FAQ ID -2496

The QIAsymphony PAXgene Blood RNA Protocol uses 2.5 ml whole human blood collected in PAXgene Blood RNA Tubes.

FAQ ID -2981

The QIAsymphony PAXgene Blood RNA Kit can be used for 96 RNA isolations.

FAQ ID -2497