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PAXgene Bone Marrow RNA Kit

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

  • 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
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.
Cat No./ID: 764133
PAXgene Bone Marrow RNA Kit (30)
€536.00
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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
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.
0
Stability of 5 transcripts at room temperature.
The changes in mRNA levels (ΔCT) from day 0 to day 2 for the samples in figure "RNA stability at room temperature" were calculated for each transcript quantified. The minimal changes and small standard deviation for samples stabilized in PAXgene Bone Marrow RNA Tubes contrasts with the significant changes for samples stored in EDTA tubes.
1
RNA yields from bone marrow samples.
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.
2
RNA stabilized at 2–8°C.

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.

3
High RNA integrity after freeze–thaw cycles.

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 up to 2 freeze–thaw cycles.

4
RNA stability at room temperature.
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.
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Storage under different conditions.
Bone marrow samples were stored in PAXgene Bone Marrow RNA Tubes [A] for 3 days at 18–25°C, [B] for 5 days at 2–8°C, or [C] subjected to 2 freeze–thaw cycles. RNA was purified using the PAXgene Bone Marrow RNA Kit, and RNA purity was analyzed by absorbance. The A260/A280 ratios indicate highly pure RNA from samples stored under different conditions.
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").
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
Features
Specifications
Applications RT-PCR, array analysis
Elution volume 40 µl
Format Spin column
Main sample type Bone marrow samples
Processing Manual (centrifugation)
Sample amount 2 ml
Technology Silica technology
Time per run 95 min/12 samples
Yield 11 µg

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

 


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