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 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.](/storage/images/_processed_/1/a/csm_RNA_stability_at_room_temperature_346d5eb789.jpg "Figure 5. RNA stability at room temperature.")
