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QIAsymphony PAXgene Blood RNA Kit

For automated purification of intracellular RNA (including miRNA) from whole blood

  • Optimized protocol enables 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
Feature

Specification

Format  

Magnetic beads

Technology

Magnetic particle technology

Sample amount

2.5 ml

Elution volume

80 µl or 120 µl or 200 µl

Main sample type 

Whole blood

Time per run  

2.5 h for 24 samples

Typical RNA yield

8–16 µg/2.5 ml sample

RNA purity 

1.8–2.2 (A260/A280)

Processing

Automated: QIAsymphony SP


Intended Use

The QIAsymphony PAXgene Blood RNA Kit is intended for the purification of RNA from whole blood collected in PAXgene Blood RNA Tubes and processed on the QIAsymphony SP. The QIAsymphony PAXgene Blood RNA Kit is intended for purification of intracellular RNA, including miRNA, from human whole blood. The QIAsymphony PAXgene Blood RNA Kit is not intended for the isolation of genomic DNA or viral nucleic acids from human whole blood.

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.

Order information

Product

Catalog Nr.

Price

QIAsymphony PAXgene Blood RNA Kit (96)

For 96 preps: 2 Reagent Cartridges, Enzyme Racks, accessories, and RNase-free buffers. To be used in conjunction with PAXgene Blood RNA Tubes.

762635
1071 USD
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Details

Performance

The fully automated QIAsymphony 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 QIAsymphony PAXgene Blood RNA procedure provides high-quality RNA from different donor samples with enrichment of small RNA species (Figure 1). Since yields are highly donor-dependent, individual yields may vary. The integrated DNase treatment removes trace genomic DNA and on-board measures of the QIAsymphony SP minimize cross-contamination (Figure 2).

Principle

Copy numbers of individual RNA species in blood can change significantly during storage or transport at ambient temperature, making reliable studies of gene expression difficult. Both stabilization of RNA molecules after collection and an efficient RNA purification protocol are needed to maximize insights into expression profiles of whole blood. Based on proven magnetic particle technology, and when used in combination with PAXgene Blood RNA Tubes, the QIAsymphony PAXgene Blood RNA Kit provides a rapid, automated procedure to isolate intracellular RNA, including miRNA, of high quality and purity.

Procedure

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 (Figure 3). 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 (Figure 2). 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.

Applications

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

  • cDNA synthesis
  • Gene expression arrays
  • End-point RT-PCR
  • Quantitative RT-PCR, including TaqMan technology
  • Next-generation sequencing
Blood was collected into PAXgene Blood RNA Tubes. RNA was purified using the QIAsymphony PAXgene Blood RNA Kit on the QIAsymphony SP instrument. RNA was analyzed with an Agilent 2100 Bioanalyzer. The arrows indicate small RNA species (including miRNA). RIN: RNA integrity number.Zoom image

Figure 1. High quality RNA from different donors.

Blood was collected into PAXgene Blood RNA Tubes. RNA was purified using the ...
Tip guards minimize the risk of sample-to-sample cross-contamination. [A] Pipetting channels. [B] Extractor unit.Zoom image

Figure 2. On-board measures against cross-contamination.

Tip guards minimize the risk of sample-to-sample cross-contamination. [A] ...
[A] QIAsymphony SP instrument. [B] Schematic of the QIAsymphony SP principle.Zoom image

Figure 3. QIAsymphony PAXgene Blood RNA procedure.

[A] QIAsymphony SP instrument. [B] Schematic of the QIAsymphony SP principle.  ...

Resources

QIAsymphony PAXgene Blood RNA Kit


1. Which sample types can be processed using the QIAsymphony PAXgene Blood RNA protocol?
2.5 ml whole human blood collected in PAXgene Blood RNA Tubes.

2. Do the samples require pretreatment before loading onto the instrument?
Yes. Samples 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 has 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).

3. What are the expected yields of RNA from PAXgene Blood RNA Tubes using the QIAsymphony PAXgene Blood RNA Kit?
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 QIAsymphony PAXgene Blood RNA System). Since yields are highly donor-dependent, individual yields may vary. Mean yields are in the range of 8–10 µg per tube or higher.

4. How should the samples be stored before purification of RNA?
Room temperature storage of the filled PAXgene Blood RNA Tubes for up to 3 days may have no effect on the RNA profile. For longer storage, they should be stored at –20°C or –70°C.

5. How many samples can be processed with the QIAsymphony PAXgene Blood RNA Kit?
The kit can be used for 96 RNA preps.

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

7. What elution formats can be used for the QIAsymphony PAXgene Blood RNA protocol?
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.

8. Why is the final heat denaturation step so important?
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.

9. Is it possible to isolate genomic DNA or viral nucleic acids with the QIAsymphony PAXgene Blood RNA protocol?
The product is not for the isolation of genomic DNA or viral nucleic acids from human whole blood.

10. Can the QIAsymphony PAXgene Blood RNA system be used for diagnostic or prognostic procedures?
No. The product 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.

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