PAXgene Blood miRNA Kit for microRNA Isolation

For purification of miRNA and total RNA from whole blood

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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

ZAR 13,610.00

Quantity

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.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

Effective isolation of intracellular RNA, including miRNA
Standardized sample processing prior to analysis
Integrated DNase treatment for removal of gDNA
Isolation can be automated on QIAcube instruments

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

The protocols of the PAXgene Blood miRNA Kit provide optimal binding conditions for several RNA species. The total RNA isolated includes mRNA and RNA molecules smaller than 200 nucleotides, such as 5.8S rRNA, 5S rRNA, tRNAs, and miRNAs (see figure "High yields of small RNA species"). The chemistry of the PAXgene Blood miRNA Kit is complimentary to various downstream applications including QIAGEN's PCR technologies. The obtained high quality miRNA can be analyzed using the QIAGEN miScript PCR System* (see figure "Analysis of miRNA species") or miRCURY LNA miRNA Probe PCR System (see figure "analysis of miRNA using miRCURY LNA miRNA Probe PCR Assays with miRCURY LNA miRNA Probe PCR System").

*This product is discontinued and my not be available for purchase in your region.

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 isolation protocol are needed to maximize insights into expression profiles of whole blood. Based on proven silica-membrane technology in trusted spin column format, and when combined with PAXgene Blood RNA Tubes, the PAXgene Blood miRNA Kit provides the chemistry and rapid procedure to isolate high-quality total RNA and miRNA.

Procedure

Cell lysis and stabilization of intracellular RNA takes place upon collection of whole blood in PAXgene Blood RNA Tubes, effectively preventing changes to gene expression profiles by ex vivo gene regulation or RNA degradation. After removal of cellular debris, kit buffers provide optimal conditions for RNA molecules to bind to the silica membrane. Contaminants are washed away and trace DNA is removed by treatment with DNase I. Pure RNA, including miRNA, is then eluted in a low-salt elution buffer and denatured by heating (see figure "PAXgene Blood miRNA procedure").

Applications

The isolated RNA 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

Supporting data and figures

Analysis of miRNA species.

Blood was collected in PAXgene Blood RNA Tubes and stored for 20–24 hours at room temperature before freezing at –15 to–30°C. RNA was purified using the PAXgene Blood RNA Kit or the PAXgene Blood miRNA Kit, as indicated. miRNA species were analyzed using the QIAGEN miScript PCR System, with the indicated miScript Primer Assays. The lower C_T values for RNA purified using the PAXgene Blood miRNA Kit indicate higher amounts of those miRNA species.

Analysis of miRNA using miRCURY LNA miRNA Probe PCR Assays with miRCURY LNA miRNA Probe PCR System

Specifications

Features	Specifications
Applications	RT-PCR and real-time RT-PCR, differential display, cDNA synthesis, northern, dot, and slot blot analyses, primer extension, microarray analysis
Elution volume	80 µl
Time per run or per prep	Manual: 90 min/12 samples; Automated: 125 min/12 samples, Reduced hands-on time.
Technology	Silica technology
Main sample type	Whole blood
Yield	3–18 µg/2.5 ml sample
Format	Spin column
Processing	Manual (centrifugation) or automated on QIAcube instruments
Sample amount	2.5 ml

Resources

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

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

Voss et al., AMP 2009

Voss et al., qPCR 2009

Kruhoffer et al., AMP 2007

This note is to inform you that the street address for PreAnalytiX GmbH has changed from “Feldbachstrasse” to “Garstligweg 8”. Please be informed that the update of the product labeling to the new address is ongoing.

In order to reduce paper consumption and oblige the growing number of customers requesting an environmentally friendly alternative to traditionally printed handbooks, we are now providing kit handbooks for Research Use Only (RUO) PreAnalytiX kits on our website only.

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

For purification of miRNA and total RNA from whole blood.

FAQ

You can find additional information relating to the PreAnalytiX PAXgene products on the PreAnalytiX website .

FAQ ID - 3515

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

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

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

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

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

FAQ ID - 3482

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

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

2.5 ml whole human blood collected in PAXgene Blood RNA Tubes

FAQ ID - 3478

Stability data are not yet available, but a stability study was started.

FAQ ID - 3480

The kit can be used for 50 RNA isolations.

FAQ ID - 3477

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