PAXgene Blood miRNA Kit

For purification of miRNA and total RNA from whole blood

Features

  • Integrated system for collection, stabilization, and purification
  • Effective purification of miRNA and total RNA
  • RNA stabilization for up to 3 days at 18–25°C
  • Stabilization for at least 50 months at –20°C or –70°C
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PAXgene Blood miRNA Kit (50)

Cat. No. / ID: 763134

For 50 RNA preps: PAXgene Spin Columns, PAXgene Shredder Spin Columns, Processing Tubes, Microcentrifuge Tubes, RNase-Free DNase, RNase-Free Reagents and Buffers; to be used in conjunction with PAXgene Blood RNA Tubes
$738.00
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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.

Product Details

The PAXgene Blood miRNA System consists of the PAXgene Blood miRNA Kit for purification of total RNA, including RNA longer than approximately 18 nucleotides, and PAXgene Blood RNA Tubes (available from BD, cat. no. 762165) for collection, stabilization, and transport of blood. Purification is carried out using silica-based RNA purification technology in a spin-column format. Purification can be carried out manually, using a microcentrifuge, or automated on the QIAcube Connect. Used with the tubes, the kit provides a complete preanalytical solution for collection and stabilization through to purification of high-quality total RNA, including miRNA, for molecular analysis.

Performance

Blood samples collected in PAXgene Blood RNA Tubes can be safely stored or transported at 15–25°C for up to 72 hours, at 2–8°C for up to 5 days, or at –20°C or –70°C for at  least 50 months without showing any significant RNA degradation or changes in transcript levels. Total RNA >18 nucleotides is purified from the stabilized blood samples with the PAXgene Blood miRNA Kit and includes both mRNA and small RNAs such as miRNA (see figure " High yields of small RNA species"). The miRNA species can be analyzed using the QIAGEN miScript PCR System (see figure " Analysis of miRNA species") or other real-time RT-PCR systems.
See figures

Principle

Blood samples are collected in PAXgene Blood RNA Tubes. The tubes contain a reagent that lyses blood cells and immediately stabilizes intracellular RNA to preserve the gene expression profile. RNA stabilization is critical for reliable downstream gene expression analysis. Without stabilization, degradation of RNA and upregulation or downregulation of transcripts occur immediately after blood is drawn.

Procedure

With the PAXgene Blood miRNA Kit, total RNA >18 nucleotides (including miRNA) is purified from the stabilized blood samples using well-established PAXgene silica-membrane technology. Purification can be carried out in a manual format or it is automatable on the QIAcube (see figure " The QIAcube").
See figures

Applications

The purified total RNA is ready to use and is ideally suited for any downstream application, including:

  • RT-PCR and real-time RT-PCR
  • Differential display
  • cDNA synthesis
  • Northern, dot, and slot blot analyses
  • Primer extension
  • Poly A+ RNA selection
  • RNase/S1 nuclease protection
  • Microarray analysis

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsRT-PCR and real-time RT-PCR, differential display, cDNA synthesis, northern, dot, and slot blot analyses, primer extension, microarray analysis
Elution volume40 µl
Time per run or per prep90 minutes/12 samples
TechnologySilica technology
Main sample typeWhole blood
Yield>3 µg (total RNA)
FormatSpin column
ProcessingManual (centrifugation) or on QIAcube
Sample amount2.5 ml

Resources

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

 

For purification of intracellular RNA, including miRNA, from whole blood

 

Application Notes (2)
Comparison of miRNA recovery from the PAXgene Blood miRNA System and the Tempus Blood RNA Tube processed with the Tempus Spin RNA Isolation Kit

 

Typical total RNA yields from PAXgene Blood RNA Tubes processed with the PAXgene Blood miRNA Kit
Kit Handbooks (1)
For purification of miRNA and total RNA from whole blood.

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 miRNA System be used for diagnostic or prognostic procedures?
No. The PAXgene Blood miRNA Kit 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.
FAQ ID -2494
What are the expected yields of RNA from PAXgene Blood RNA Tubes isolated with the PAXgene Blood miRNA kit?
Typical yields of RNA isolated from 2.5 ml healthy, human, whole blood (4.8 x 106 –1.1 x 107 leukocytes/ml) are ≥3 μg for >95% of the samples processed (Refer to Technical Note) . Since yields are highly donor-dependent, individual yields may vary.
FAQ ID - 3481
Is it possible to isolate genomic DNA or viral nucleic acids with PAXgene Blood miRNA protocol?
No. The PAXgene Blood RNA System has been optimized for cellular RNA only. PreAnalytiX offers a dedicated system for collection and isolation of genomic DNA from human whole blood (see PAXgene Blood DNA System) .
FAQ ID - 3483
What can I use to isolate RNA smaller than 200 nucleotides?

For the isolation of microRNA (miRNA) specifically, we have developed the miRNeasy Mini Kit and the miRNeasy 96 Kit for isolation from cells and tissues.   We also have the PAXgene Blood miRNA and Tissue miRNA kits for isolation from blood stored in PAXgene Blood RNA tubes and PAXgene Tissue Containers, respectively.  Other miRNA isolation supplementary protocols can also be found by searching our comprehensive protocols at http://www.qiagen.com/literature/default.aspx?WT.svl=m.

FAQ ID -115
What is the maximum number of samples for the QIAcube variant of the PAXgene Blood miRNA protocol for a single run?

Up to 12 samples can be processed in a single run.

FAQ ID - 3482
How great is the enrichment of miRNA when using the dedicated PAXgene Blood miRNA Kit compared to the PAXgene Blood RNA Kit?

If the PAXgene Blood miRNA Kit is used, an enrichment of small RNAs as part of the total RNA is achieved. The enrichment may vary between different miRNA species and samples, but on average an enrichment of about three or more CT-values can be expected.

FAQ ID -2495
Do eluates generated with the PAXgene Blood miRNA Kit contain residual genomic DNA?
Typically, the amount of genomic DNA (gDNA) in eluates of the PAXgene Blood miRNA Kit is very low as long as the kit is stored and used according to the instructions of the PAXgene Blood miRNA Kit Handbook. For downstream applications that are especially sensitive to gDNA contamination, a specialized protocol for the QIAcube helps reduce gDNA content further and can be downloaded from the QIAcube product page.
FAQ ID - 3637
Which sample types can be processed using the PAXgene Blood miRNA protocols?
2.5 ml whole human blood collected in PAXgene Blood RNA Tubes
FAQ ID - 3478
Do you have stability data for RNA isolated with PAXgene miRNA Kit?
Stability data are not yet available, but a stability study was started.
FAQ ID - 3480
How many samples can be processed with the PAXgene Blood miRNA Kit?
The kit can be used for 50 RNA isolations.
FAQ ID - 3477
Do the samples require pretreatment before starting the manual protocol or before loading onto the QIAcube instrument?
Yes. Samples have to be incubated at least 2 h 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 three days has no effect on the RNA profile. During the preparation of the RNA the manual pretreatment consist of a centrifugation and a pellet resuspenion step (see the PAXgene Blood miRNA Handbook ).
FAQ ID - 3479