PAXgene Blood RNA Kit (IVD)

For purification of intracellular RNA from whole blood to be used for in vitro diagnostic (IVD) tests

S_1391_RPA_PAX1163

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PAXgene Blood RNA Kit (50)

Cat. No. / ID: 762174

50 PAXgene Spin Columns, 50 PAXgene Shredder Spin Columns, Processing Tubes, RNase-Free DNase I, RNase-Free Reagents and Buffers. To be used in conjunction with PAXgene Blood RNA Tubes
SEK 7,290.00
These products are not available in all countries; please inquire.
The PAXgene Blood RNA Kit (IVD) is intended for in vitro diagnostic use.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • For in vitro diagnostic use

  • Coupled with the PAXgene Blood RNA Tube (IVD), constitutes the PAXgene Blood RNA System

  • Efficient purification of high quality intracellular RNA

  • Standardized sample processing prior to analysis

  • Integrated DNase treatment for the removal of gDNA

  • Purification can be automated on the QIAcube Connect MDx*

*The QIAGEN QIAcube Connect MDx is not available in all countries. For further details please contact QIAGEN Technical Service.

Product Details

The PAXgene Blood RNA System consists of a blood collection tube (PAXgene Blood RNA Tube) and a nucleic acid purification kit (PAXgene Blood RNA Kit). It is intended for the collection, storage and transport of blood and stabilization of intracellular RNA in a closed tube, and subsequent isolation and purification of host RNA from whole blood for RT-PCR used in molecular diagnostic testing.

Performance characteristics for the PAXgene Blood RNA System have only been established with FOS* and IL1B** gene transcripts. The user is responsible for establishing appropriate PAXgene Blood RNA System performance characteristics for other target transcripts.

*FOS is the gene symbol for human Fos Proto-Oncogene, AP-1 Transcription Factor Subunit.

**IL1B is the gene symbol for human Interleukin 1 Beta.

Performance

Total RNA purified using the PAXgene Blood RNA System is highly pure, with A260/A280 values between 1.8 and 2.2 and ≤1.0% (w/w) genomic DNA in ≥95% of all samples processed (see "RNA yield, purity and genomic DNA contamination - automated processing."). At least 95% of samples show no inhibition in RT-PCR, when eluate accounts for up to 30% of the RT-PCR reaction. RNA yields from 2.5 ml healthy human whole blood are ≥3 µg for ≥95% of the samples processed. Since yields are highly donor-dependent, individual yields may vary. For individual donors, the PAXgene Blood RNA System provides highly reproducible and repeatable yields (see figures " Reproducible and repeatable RNA purification" and " Repeatability and reproducibility of RNA yield"), making the system highly robust for clinical diagnostic tests.

The automated protocol of RNA purification using the PAXgene Blood RNA System provides pure RNA in high yield with reproducible and repeatable RT-PCR results, as shown in the figures (see "RNA yield, purity and genomic DNA contamination - automated processing.").

See figures

Principle

Copy numbers of individual RNA species in blood can change significantly during sample 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 silica-membrane technology in spin column format, and when combined with PAXgene Blood RNA Tubes, the PAXgene Blood RNA Kit provides a rapid procedure and the chemistry to isolate RNA of high quality and purity.

Procedure

The PAXgene Blood RNA procedure is simple and can be performed using manual or automated procedures (see flowcharts " Manual PAXgene Blood RNA procedure" and " Automated PAXgene Blood RNA procedure").

Manual PAXgene Blood RNA procedure

The PAXgene Blood RNA procedure begins with a centrifugation step to pellet nucleic acids in the PAXgene Blood RNA Tube. The pellet is washed, resuspended, and incubated in optimized buffers together with proteinase K to bring about protein digestion. An additional centrifugation through the PAXgene Shredder spin column is carried out to homogenize the cell lysate and remove residual cell debris, and the supernatant of the flow-through fraction is transferred to a fresh microcentrifuge tube. Ethanol is added to adjust binding conditions, and the lysate is applied to a PAXgene RNA spin column. During a brief centrifugation, RNA is selectively bound to the PAXgene silica membrane as contaminants pass through. Remaining contaminants are removed in several efficient wash steps. Between the first and second wash steps, the membrane is treated with DNase I to remove trace amounts of bound DNA. After the wash steps, RNA is eluted in elution buffer and heat-denatured.

Automated PAXgene Blood RNA procedure

Sample preparation, automated on the QIAcube Connect MDx, follows the same steps as the manual procedure, enabling you to continue using the PAXgene Blood RNA Kit for purification of high quality RNA.

The procedure begins with a centrifugation step to pellet nucleic acids in the PAXgene Blood RNA Tube. The pellet is washed, resuspended, and transferred from the PAXgene Blood RNA Tube into processing tubes, which are placed into the thermoshaker unit on the QIAcube Connect MDx worktable. The operator selects and starts the "PAXgene Blood RNA Part A" protocol from the menu. The QIAcube Connect MDx performs the steps of the protocol through to elution of RNA in elution buffer. The operator transfers the microcentrifuge tubes, containing the purified RNA, into the thermoshaker unit of the QIAcube Connect MDx. The operator selects and starts the "PAXgene Blood RNA Part B" protocol from the menu, and heat denaturation is performed by the QIAcube Connect MDx. Average sample preparation time (based on data for 12 sample preps) is 151 minutes, with only approximately 20 minutes of hands-on time.

See figures

Applications

When the kit is used in conjunction with PAXgene Blood RNA Tubes, the system provides intracellular RNA from whole blood for RT-PCR used in molecular diagnostic testing.

Supporting data and figures

Specifications

FeaturesSpecifications
FormatSpin column
TechnologySilica technology
Sample amount2.5 ml
Elution volume80 µl (2 x 40 µl)
Main sample typeWhole blood
Time per runManual: 90 min/12 samples; Automated: 151 min/12 samples
Yield≥3 µg/2.5 ml sample (≥95% of all samples processed). Apparently healthy consented donors with white blood cell counts in range of 4.8 x 10^6 – 1.1 x 10^7 leukocytes/ml.
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein1.18-2.2 (A260/A280) RNA; ≤1 % (w/w) genomic DNA (≥95% of all samples processed)
ProcessingManual: centrifugation; Automated: QIAcube Connect MDx

Resources

Kit Handbooks (1)
Brochures & Guides (3)
For collection, transport, and storage of whole blood and stabilization and purification of intracellular RNA

 

Safety Data Sheets (1)
Download Safety Data Sheets for QIAGEN product components.
Protocol Files (1)
Application Notes (2)
In situ stability of RNA in blood specimens stored for 11 years (132 months) at –20°C and –70°C in PAXgene Blood RNA Tubes

 

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

FAQ

Where can I find additional information for PreAnalytiX PAXgene products?
You can find additional information relating to the PreAnalytiX PAXgene products on the PreAnalytiX website .
FAQ ID - 3515
Can the PAXgene Blood RNA System be used with animal blood samples?

The PAXgene Blood RNA System was developed for intracellular RNA isolation from human blood samples. QIAGEN offers RNA stabilization and isolation systems for animal blood. Please contact QIAGEN technical service (www.qiagen.com) for more information.

FAQ ID - 3461
Can the PAXgene Blood RNA Tubes undergo freeze/thaw cycles?
PAXgene Blood RNA Tubes can go through up to two freeze/thaw cycles with no impact on yield or mRNA stability.
FAQ ID -2471
What RNA yields are expected with the PAXgene Blood RNA kit?

Typical yields of RNA isolated from 2.5 ml human whole blood are between 4 and 20 µg. However yield is highly donor-dependent, and in some cases higher or lower yields may be achieved. At least 95% of all samples with a WBC count between 4.8 and 11.0 x 106 cells/ml of blood will yield ≥3 µg of total RNA per tube. To prevent low yields due to underfilling of the tube, see the phlebotomy FAQ and the blood collection demonstration video.

FAQ ID - 3464
Why is a blood collection set required to be used with the PAXgene Blood RNA tube?

A blood collection set is required to be used with the PAXgene Blood RNA Tube to eliminate the possibility of patient contact with the reagent in the tube, due to backflow of blood, when used in accordance with the instructions for use.

FAQ ID - 3459
What is the integrity of RNA from PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue?
Like yield, RNA integrity depends on parameters, such as tissue type, period of cold ischemia, fixation time, processing protocol, age, and storage conditions of the PFPE block. For examples of the integrity of RNA in rat tissues under ideal workflow conditions, see Groelz et al., Exp Mol Pathol. 2013 Feb;94(1). For examples of the integrity of RNA from clinical samples, see Viertler et al., J Mol Diagn. 2012 Sep;14(5)
FAQ ID -3046
How long can the PAXgene Blood RNA tubes be kept frozen at –20°C or –70°C?
Currently we have data for storage at –20°C and –70°C for 96 months (In situ stability of RNA in blood specimens stored for 8 years (96 months) at –20°C and –70°C in PAXgene Blood RNA Tubes).  

Long term archiving studies are ongoing. See technical note: In Situ Stability of RNA in Blood Samples Stored at –20°C and –70°C in PAXgene Blood RNA Tubes.


 
FAQ ID - 3463
What are possible reasons for blood draws with lower than expected blood volume?

Possible reasons for blood draws with lower than expected blood volume include: 1) The phlebotomist has not waited enough time for the blood to stop flowing into the tube. 2) The tube has "pushed back" from the needle end attached to the holder and the tube is no longer drawing. 3) Discard tube was not used. For further information see the video Collecting Specimens In PAXgene Blood RNA Tubes and Instructions for Use.

FAQ ID - 3462
Is there any chance of the PAXgene Blood RNA tube reagent going back into the patient's arm?

If used according to the instructions for use, there is no possibility of backflow during phlebotomy, and thus no possibility of reagent going into the patient’s arm.

FAQ ID - 3460