PAXgene Bone Marrow RNA Kit

For collection of bone marrow samples and stabilization/purification of intracellular RNA

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PAXgene Bone Marrow RNA Kit (30)

Cat. No. / ID: 764133

For 30 RNA preps: 30 PAXgene Spin Columns, 30 PAXgene Shredder Spin Columns, Processing Tubes, RNase-free DNase I, RNase-free reagents and buffers. To be used in conjunction with PAXgene Bone Marrow RNA Tubes

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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.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

To be used in conjunction with the PAXgene Bone Marrow RNA Tube
Rapid purification of RNA with high integrity and purity
Optimized protocol for heterogeneous bone marrow samples
Integrated DNase treatment for gDNA removal

Product Details

The PAXgene Bone Marrow RNA System consists of PAXgene Bone Marrow RNA Tubes, for stabilization, storage, and transportation of bone marrow, and the PAXgene Bone Marrow RNA Kit, for silica-membrane-based RNA isolation and purification in a spin-column format. Bone marrow samples are collected using standard medical practices, with commonly used anticoagulants. A 2 ml sample is immediately transferred into a PAXgene Bone Marrow RNA Tube, which stabilizes the RNA in the sample for storage at room temperature, refrigerated, or frozen. RNA can be purified days later using the PAXgene Bone Marrow RNA Kit or archived for several months at –20°C or –70°C.

Performance

The PAXgene Bone Marrow RNA Kit enables efficient isolation and purification of intracellular RNA from heterogeneous human whole bone marrow samples (see figure ""), including an optimized procedure for cell-rich, viscous samples. RNA can be purified days after collection, and intact, high-quality RNA can be purified using the PAXgene Bone Marrow RNA Kit after samples are stored at room temperature, refrigerated, or frozen (see figures "", "", and ""). RNA Tubes protect RNA molecules from degradation by RNases and enable accurate analysis by minimizing ex vivo changes in gene expression (see figure ""). An integrated DNase treatment ensures that genomic DNA is minimized. In-house and external studies showed that genomic DNA was <0.85% in 95% of the samples tested (n = 157). A₂₆₀/A₂₈₀ values are typically between 1.8 and 2.2, indicating the high purity of the RNA (see figure "").

See figures

RNA yields from bone marrow samples.

RNA stability at room temperature.

RNA stabilized at 2–8°C.

High RNA integrity after freeze–thaw cycles.

Stability of 5 transcripts at room temperature.

Storage under different conditions.

Principle

Copy numbers of individual mRNA species in bone marrow can change significantly during storage or transport at room temperature, making reliable studies of gene expression impossible. PAXgene Bone Marrow RNA Tubes protect RNA molecules from degradation and immediately stabilize the RNA in the sample for storage at room temperature, refrigerated, or frozen.

Procedure

Bone marrow samples are collected using standard medical practices, with commonly used anticoagulants. A 2 ml sample is then transferred into a PAXgene Bone Marrow RNA Tube. The RNA in the sample is immediately stabilized for storage at room temperature, refrigerated, or frozen. The PAXgene Bone Marrow RNA Kit provides an easy and rapid procedure for isolation and purification of intracellular RNA from human whole bone marrow samples. The kit uses silica-membrane technology in a spin-column format for efficient purification of intact, high-quality RNA.

Applications

RNA purified using the PAXgene Bone Marrow RNA Kit is ready for use in a wide range of downstream applications, including

Real-time RT-PCR analysis of specific transcripts
Array analysis of complex expression profiles

Supporting data and figures

Stability of 5 transcripts at room temperature.

Specifications

Features	Specifications
Applications	RT-PCR, array analysis
Yield	11 µg
Time per run	95 min/12 samples
Main sample type	Bone marrow samples
Sample amount	2 ml
Processing	Manual (centrifugation)
Format	Spin column
Elution volume	80 µl (2 x 40 µl)
Technology	Silica technology

Resources

For collection, storage, and transport of bone marrow samples and stabilization and purification of intracellular RNA

Download Safety Data Sheets for QIAGEN product components.

Guenther et al., AMP 2006

Guenther et al., AACR 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.

For isolation and purification of intracellular RNA from whole bone
marrow samples stabilized in PAXgene Bone Marrow RNA Tubes

For the collection of human bone marrow and immediate stabilization of intracellular RNA

FAQ

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

FAQ ID - 3515

As gene expression starts to change in a sample removed from the human body within minutes, we highly recommend transferring the samples within a few minutes into the stabilization solution of the PAXgene Bone Marrow RNA tubes for cell lysis directly linked to RNA stabilization to take place.

FAQ ID -2504

Yes. Heating temperature and incubation time will vary depending on different RT protocols. Therefore, the final heating step should be performed routinely at the end of the protocol.

FAQ ID -2513

No. The elution buffer (Buffer BMR5) supplied in the PAXgene Bone Marrow RNA Kit must be used.

FAQ ID -2512

No. The cells are lysed in the PAXgene Bone Marrow RNA Tube. There is no method available to enrich for certain sub-populations of cells.

FAQ ID -2508

The supernatants resulting from the first two spin steps of the PAXgene Bone Marrow RNA Kit purification procedure may still contain biologically hazardous material. We recommend classifying the tube-processed supernatant of the kit as both a biological and chemical waste. Supernatants can be decontaminated by autoclaving and disposed of as chemical waste. Disposal should be done in accordance with local, state and federal regulations.

CAUTION: Do not add bleach or acidic solutions directly to the sample preparation waste.

FAQ ID -2516

The PAXgene Bone Marrow RNA System was developed for intracellular RNA isolation from human bone marrow samples. Bone marrow samples from other species or from blood were not tested.

FAQ ID -2501

Yes. We have tested the PAXgene Bone Marrow RNA System with bone marrow samples containing very high numbers of leukocytes. Bone marrow samples with higher leukocyte numbers are indicated by a highly viscous solution after addition of buffers BMR1 and BMR2 followed by incubation. These challenging samples are further diluted according to descriptions given in the protocol and do not lead to problems in the extraction procedure.

FAQ ID -2510

Yes. You may dispose of the waste product from the PAXgene Bone Marrow RNA Tube with a 5% sodium hypochlorite bleach solution, in a 1:9 ratio (1 part bleach : 9 parts tube contents).

FAQ ID -2515

The PAXgene Bone Marrow RNA Tubes should be frozen at –20°C and –70°C in a wire rack, not in an expanded polystyrene foam tray.

If the PAXgene Bone Marrow RNA Tubes are to be stored at temperatures lower than –20ºC, freeze them first for 24 h at –20ºC, then transfer them to –70ºC.

Currently available data shows stabilization of cellular RNA for several months at –20°C or –70°C. For more information from ongoing studies evaluating stability for longer time periods, please contact QIAGEN Technical Services.

FAQ ID -2505

Bone marrow samples are extremely heterogeneous, consisting of varying amounts of cells, tissue, and solid matter. RNA yields are highly donor-dependent and can vary greatly from sample to sample as well as between replicates from the same donor due to sample-intrinsic heterogeneity.

In a study of 295 bone marrow samples, yields of RNA isolated from 2 mL human bone marrow were between 2.0 and 44.7 µg (5-95% quantiles, mean = 15.8 µg, median = 11.0 µg).

FAQ ID -2506

Yes. The PAXgene Bone Marrow RNA System purifies RNA from whole bone marrow. Therefore it will also purify RNA from other cells, e.g. reticulocytes and RBCs. Array analyses have shown higher globin mRNA levels for RNA isolated from whole bone marrow compared to RNA isolated from mononuclear cells.

FAQ ID -2509

No. The 260/230 nm absorbance ratio is not specified for the PAXgene Bone Marrow RNA extraction kit. In contrast to a reduced 260/280 ratio, which results from a protein contamination of the eluted RNA and can lead to inhibitory effects in down stream applications, there is no negative impact of a reduced 260/230 ratio on any down stream application reported to date and therefore it is not necessary to specify this.

In addition, because the elution buffer contains very low concentrations of salt that absorb at wavelengths between 220 and 230 nm, it is necessary to zero the photometer properly. Do this by preparing a solution for zeroing the photometer which contains a similar salt concentration as the sample to be measured by using the same volumes. Add the same volume of Buffer BMR5 as the volume of RNA sample to be diluted into a fresh volume of the buffer in which you will measure the absorption (we recommend 10 mM Tris Cl pH 7.5 buffer for this purpose).

FAQ ID -2514

The PAXgene Bone Marrow RNA kit elution buffer (Buffer BMR5) is a proprietary low-salt buffer. In combination with the final heat denaturation step, it leads to optimal performance of the isolated RNA in RT-reactions. There is no interference with downstream applications reported to date.

FAQ ID -2511

No. The PAXgene Bone Marrow RNA System was developed for intracellular RNA isolation from human bone marrow samples. PreAnalytiX offers a dedicated collection and RNA extraction system for human blood (see the PAXgene Blood RNA System). QIAGEN also offers RNA stabilization and isolation systems for saliva and cultured cells. Please contact QIAGEN technical service for more information or visit www.qiagen.com.

FAQ ID -2502

No. Transferring bone marrow volumes different than 2mL into the PAXgene Bone Marrow RNA Tube is not recommended. The tube and its contents were optimized for performance with 2 mL of bone marrow. The use of other volumes is not recommended.

FAQ ID -2503

No. The PAXgene Bone Marrow RNA System has been optimized only for cellular RNA only.

FAQ ID -2507