PAXgene Bone Marrow RNA Tubes

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

Features

  • Integrated system for storage, stabilization, and purification
  • Immediate stabilization of intracellular RNA at collection
  • Rapid purification from heterogeneous bone marrow samples
  • Safe and easy transportation
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PAXgene Bone Marrow RNA Tubes (50)

Cat. No. / ID: 764114

50 Bone Marrow Collection Tubes. To be used in conjunction with the PAXgene Bone Marrow RNA Kit
₩1,537,000.00
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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 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 " RNA yields from bone marrow samples"), 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 " RNA stability at room temperature", " RNA stabilized at 2–8°C", and " High RNA integrity after freeze–thaw cycles"). RNA Tubes protect RNA molecules from degradation by RNases and enable accurate analysis by minimizing ex vivo changes in gene expression (see figure " Stability of 5 transcripts at room temperature"). 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 " Storage under different conditions").
See figures

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

Specifications

FeaturesSpecifications
Tube size16 x 125 mm
Label typePaper
Draw volume2 ml
Quantity50/box
Additive/concentrationAdditive 11.1 ml
Closure type/colorBD Hemogard/Red

Resources

Brochures & Guides (3)
For collection, storage, and transport of bone marrow samples and stabilization and purification of intracellular RNA

 

Kit Handbooks (2)
For the collection of human bone marrow and immediate stabilization of intracellular RNA
For isolation and purification of intracellular RNA from whole bone
marrow samples stabilized in PAXgene Bone Marrow RNA Tubes

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
How much time may the aspirated bone marrow sample be allowed to incubate before pipetting it into the PAXgene Bone Marrow RNA Tube?
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
I routinely heat an aliquot of the RNA eluate prior to my RT reaction. Do I still need to heat the eluate after the PAXgene Bone Marrow RNA preparation?
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
Can I use water to elute the RNA in the PAXgene Bone Marrow RNA Kit?
No. The elution buffer (Buffer BMR5) supplied in the PAXgene Bone Marrow RNA Kit must be used.
FAQ ID -2512
Can specific types of white blood cells be enriched from a PAXgene Bone Marrow RNA Tube prior to the RNA isolation?
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
How are the supernatants resulting from the first two spin steps of the PAXgene Bone Marrow RNA kit purification procedure disposed?

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
Can the PAXgene Bone Marrow RNA System be used with animal bone marrow or with blood samples?
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
Can bone marrow from leukemia patients with elevated numbers of white blood cells be processed with the PAXgene Bone Marrow RNA System?
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
Can the PAXgene RNA stabilization solution/bone marrow mixture from the PAXgene Bone Marrow RNA Tube be disposed of by adding bleach?
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
How should the PAXgene Bone Marrow RNA Tubes be frozen after collection and for how long?

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
What RNA yields are expected with the PAXgene Bone Marrow RNA System?

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
Is RNA from red blood cells also isolated when RNA is extracted from a PAXgene Bone Marrow RNA Tube?
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
Is the 260/230 nm absorbance ratio specified for RNA derived from the PAXgene Bone Marrow RNA System?

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
What is in the elution buffer in the PAXgene Bone Marrow RNA kit?
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
Can the PAXgene Bone Marrow RNA System be used for other human body fluids than bone marrow samples or cultured cells of human origin?
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
Can bone marrow aliquots other than 2 mL be pipetted into the PAXgene Bone Marrow RNA Tube?
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
Can the PAXgene Bone Marrow RNA System be used to isolate genomic DNA or viral RNA and DNA?
No. The PAXgene Bone Marrow RNA System has been optimized only for cellular RNA only.
FAQ ID -2507