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PAXgene Bone Marrow RNA Tube

For stabilization, transport and storage of intracellular RNA in bone marrow aspirate samples

  • To be used in conjuction with the PAXgene Bone Marrow RNA Kit
  • Immediate stabilization of intracellular RNA upon introducing the sample into the tube
  • Rapid purification of RNA from heterogeneous bone marrow samples using the PAXgene Bone Marrow RNA Kit
  • RNA is stable for days at room temperature or refrigerated
  • Long-term sample storage at –20°C or –70°C

Feature

Specification

Tube size

16 x 125 mm

Specimen volume needed

2 ml

Additive/concentration

Additive 11.1 ml

Length of stabilization

  • up to 3 days at room temperature (15–25°C)
  • up to 5 days at 2–8°C
  • Long-term at –20°C or –70°C

Closure type/color

BD Hemogard/Red

Label type

Paper

Quantity

50/box


Intended Use

The PAXgene Bone Marrow RNA System consists of a tube (PAXgene Bone Marrow RNA Tube) containing an RNA stabilizing additive, and a nucleic acid preparation kit (PAXgene Bone Marrow RNA Kit). It is intended for the stabilization, transport and storage of intracellular RNA in a human bone marrow aspirate (BMA) sample and the subsequent isolation and purification of total RNA.

Do not use for blood collection.

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. The performance characteristics of this product have not been fully established.

Order information

Product

Catalog Nr.

Price

PAXgene Bone Marrow RNA Tubes (50)

50 Bone Marrow Collection Tubes (2 ml). To be used in conjunction with the PAXgene Bone Marrow RNA Kit.

764114
1107 USD

Details

Performance

A unique additive in PAXgene Bone Marrow RNA Tubes protects RNA molecules from degradation by RNases and other enzymes, as demonstrated by high RNA integrity numbers (RIN) measured in samples even after 5 days of storage at 2–8°C (Figure 1). Preservation of RNA integrity is maintained even through freeze-thaw cycles (Figure 2). The additive also completely lyses all cells in the samples to halt cellular functions and minimize ex vivo changes in gene expression profiles (Figure 3).

Principle

Copy numbers of individual mRNA species in bone marrow can change significantly during storage or transport at ambient temperature, making reliable studies of gene expression impossible. A unique additive in the PAXgene Bone Marrow RNA Tubes lyses cells, halts cellular processes and protects RNA from degradation, stabilizing the RNA in the sample for storage at room temperature, refrigerated or frozen.

The PAXgene Bone Marrow RNA Tubes are optimized for use with the PAXgene Bone Marrow RNA Kit for the efficient isolation and purification of intracellular RNA from heterogeneous human whole bone marrow samples (Figure 4). RNA can be purified days after collection or the bone marrow sample can be archived for several months at –20°C or –70°C. Intact, high-quality RNA is purified from samples stored at room temperature, refrigerated or frozen (Figure 5, Figure 1, Figure 2).

Procedure

Bone marrow aspirates are collected using standard medical practice. A 2 ml sample is then transferred into a PAXgene Bone Marrow RNA Tube (Figure 6). The RNA in the sample is immediately stabilized for storage at room temperature, refrigerated or frozen. RNA can be purified using the PAXgene Bone Marrow RNA Kit.

Applications

RNA is stabilized in PAXgene Bone Marrow RNA Tubes and can be used in a range of downstream applications, including:

  • cDNA synthesis
  • Gene expression arrays
  • End-point RT-PCR
  • Quantitative RT-PCR, including TaqMan technology
Bone marrow samples were collected in duplicate and stabilized in PAXgene Bone Marrow RNA Tubes. RNA was purified from the samples using the PAXgene Bone Marrow RNA Kit with no storage or after 5 days storage at 2–8°C. RNA integrity was evaluated using the RNA integrity numbers (RIN), measured on the Agilent 2100 Bioanalyzer. Results are shown for 3 representative donors. The RIN did not change significantly during storage, indicating that the RNA remains stable in the sample.Zoom image

Figure 1. RNA stabilized at 2–8°C.

Bone marrow samples were collected in duplicate and stabilized in PAXgene Bone ...
Bone marrow samples were stabilized in PAXgene Bone Marrow RNA Tubes, and RNA was purified using the PAXgene Bone Marrow RNA Kit after the indicated number of freeze–thaw cycles. The RIN values did not change significantly, indicating that RNA remained intact in the samples for up to 2 freeze–thaw cycles.Zoom image

Figure 2. High RNA integrity after freeze-thaw cycles.

Bone marrow samples were stabilized in PAXgene Bone Marrow RNA Tubes, and RNA was ...
Bone marrow samples were collected from 39 individuals and immediately transferred and stored in PAXgene Bone Marrow RNA Tubes or EDTA tubes. RNA was purified from the samples on day 0 and day 2 using the PAXgene Bone Marrow RNA Kit. Changes in mRNA levels (ΔCT) from day 0 to day 2 were quantified for each of 5 transcripts. The minimal changes and small standard deviation for samples stabilized in PAXgene Bone Marrow RNA Tubes contrast with the significant changes for samples stored in EDTA tubes.Zoom image

Figure 3. Stability of 5 transcripts at room temperature.

Bone marrow samples were collected from 39 individuals and immediately transferred ...
Bone marrow samples were stabilized in PAXgene Bone Marrow RNA Tubes, and RNA was purified using the PAXgene Bone Marrow RNA Kit. Yields are indicated for 295 samples, from both in-house and external studies. The median yield for the 295 samples was 11.0 μg RNA per 2 ml sample. Note that RNA yields can vary greatly due to the extreme heterogeneity of bone marrow samples.Zoom image

Figure 4. RNA yields from bone marrow samples.

Bone marrow samples were stabilized in PAXgene Bone Marrow RNA Tubes, and RNA was ...
Bone marrow samples were collected from 39 individuals and immediately transferred to [A] PAXgene Bone Marrow RNA Tubes or [B] EDTA tubes. RNA was purified using the PAXgene Bone Marrow RNA Kit or a reference silica-membrane-based method at day 0 or after 2 days storage at room temperature. Expression levels of 5 marker transcripts for each sample (IL-8, p53, cox-2, IL-1β, c-fos) were quantified using real-time RT-PCR. The high correlation of CT values (threshold cycles) for the samples stabilized in PAXgene Bone Marrow RNA Tubes indicates that the levels of these transcripts did not significantly change during storage, in contrast to the samples stored in EDTA tubes.Zoom image

Figure 5. RNA stability at room temperature.

Bone marrow samples were collected from 39 individuals and immediately transferred ...
Zoom image

Figure 6. PAXgene Bone Marrow RNA procedure.

Resources

PAXgene Bone Marrow RNA Tube

Sample Handling

1. Can the PAXgene Bone Marrow RNA Tube be used for diagnostic or prognostic procedures?
No. The product is ‘For Research Use.’ Not for use in diagnostic procedures. No claim or representation is intended to provide information for the diagnosis, prevention or treatment of a disease.

2. 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.

3. Can the PAXgene Bone Marrow RNA System be used for human body fluids other 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 offers RNA stabilization and isolation systems for saliva and cultured cells. Please contact QIAGEN technical service (www.qiagen.com) for more information.

4. Can bone marrow aliquots other than 2 ml be pipetted into the PAXgene Bone Marrow RNA Tube?
No. Transferring bone marrow samples either larger or smaller than 2 ml 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 aspirate.

5. How much time may the aspirated bone marrow sample be allowed to incubate before pipetting it into the PAXgene Bone Marrow RNA Tube?
Transcript levels begin to change in as few as 10 minutes after a sample is removed from the human body. We highly recommend transferring the sample into the PAXgene Bone Marrow RNA Tube and mixing with the stabilization solution as soon as possible after collection in a syringe.

Transport and Storage

1. Can bone marrow samples be frozen directly in PAXgene Bone Marrow RNA Tubes at –70°C?
No. If samples in 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.

2. How should the PAXgene Bone Marrow RNA Tubes be frozen after collection?
Samples in 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 PAXgene Bone Marrow RNA Tubes are to be stored at temperatures lower than –20ºC, it is recommended to freeze them first for 24 h at –20ºC, and then transfer them to –70ºC.

3. How long can the tubes be kept frozen at –20°C or –70°C?
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 service (www.qiagen.com).

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