Performance
The patented additive in PAXgene Blood RNA Tubes protects RNA molecules from degradation by RNases and other enzymes and triggers the complete lysis of all cells in the sample to minimize ex vivo changes in gene expression due to gene induction and downregulation. The result is immediate and long-term stabilization of RNA in specimens, even after being stored for up to 3 days at room temperature (Figure 1 and Figure 2) or up to 5 days refrigerated (Figure 3 and Figure 4). Currently available data show stabilization of intracellular RNA for up to 11 years at –20°C or –70°C (see Technical Notes).
Principle
The collection of whole blood is the first step in many molecular assays used to study intracellular RNA. A major challenge in this type of testing is the instability of intracellular RNA, which rapidly degrades within hours after blood collection. Furthermore, certain transcript levels change after blood collection due to ex vivo gene regulation, such as induction and repression. Both RNA degradation and gene regulation can lead to an under- or overestimate of in vivo gene transcript levels.
The PAXgene Blood RNA Tube contains an additive that stabilizes the in vivo gene expression profile by reducing post-collection RNA degradation and minimizing gene induction and repression. When used in conjunction with one of the PAXgene Blood RNA Kits, the PAXgene Blood RNA Tube provides pure RNA for accurate detection and quantification of gene expression.
Procedure
Whole blood is collected directly into the PAXgene Blood RNA Tube, using standard phlebotomy practice and equipment. The RNA in the specimen is immediately stabilized for storage at room temperature, refrigerated, or frozen. RNA can be purified using one of the PAXgene Blood RNA Kits (choose the right PAXgene Blood RNA Kit). The ease of collection, stabilization of RNA at different temperatures, and elimination of additional processing steps (e.g., centrifugation, storage on ice) standardizes the workflow from collection to purification, reducing variability that can be introduced by different operators (Figure 5).
Applications
When used in conjunction with the PAXgene Blood RNA Kit, the PAXgene Blood RNA Tubes enable purification of intracellular RNA from whole blood for applications in molecular diagnostic testing, such as real-time RT-PCR analysis of specific transcripts.
PAXgene Blood RNA Tubes are intended for in vitro diagnostic testing (IVD) and are to be used in conjunction with the PAXgene Blood RNA Kit.