The PAXgene Blood miRNA System consists of the PAXgene Blood RNA Tube and PAXgene Blood miRNA Kit. The system is optimized to streamline the purification of total intracellular RNA, including small RNA species. By minimizing the unpredictability associated with RNA processing, the system provides enhanced accuracy in the analysis of total intracellular RNA (microRNA or miRNA, mRNA and other RNA species).
Purification of total RNA, including small RNAs, using the PAXgene Blood miRNA System can be performed manually or automated on the QIAcube (see protocols) and the resulting high-quality RNA is optimal for a range of downstream applications, including RT-PCR, expression-array and expression-chip analysis, and cDNA synthesis. To accommodate medium throughput needs, equivalent results are obtained by combining PAXgene Blood RNA Tubes with the QIAsymphony PAXgene Blood RNA Kit.
After efficient RNA stabilization, the PAXgene Blood miRNA System is designed to enrich RNA molecules of small molecular weight, such as miRNA, in conjunction with larger intracellular RNA. By maximizing their yield, the PAXgene Blood miRNA System facilitates profiling miRNA, mRNA and other RNA species.
The unique additive in the PAXgene Blood RNA tube triggers a series of stabilizing events at the time of blood collection that safeguard RNA molecules from changes caused by ex vivo gene induction and downregulation, initiation of cellular processes like programmed cell death, and enzymatic activity. Subsequent purification procedures enrich smaller RNA species, such as miRNA, for their optimal profiling.
The additive in PAXgene Blood RNA Tubes stabilizes intracellular RNA in collected specimens, enabling shipping, processing and storage at room temperature (15–25°C), refrigerated (2–8°C) or frozen (–20°C or –70°C). Frozen specimens are suitable for long-term storage without impacting the intracellular RNA profile and the integrity of subsequently purified RNAs. The PAXgene Blood miRNA System provides users with a completely integrated approach to RNA purification – from blood collection to pure RNA.
The PAXgene Blood miRNA Kit uses a unique version of QIAGEN’s proven separation and purification technology. It systematically digests proteins, filters out cellular debris and depletes genomic DNA through a DNase treatment prior to eluting RNA molecules bound to a silica membrane. Optimized chemistry enhances binding of RNA to the silica membrane, including miRNAs and other small RNA species, from samples that have been stabilized in the PreAnalytiX reagent, thereby leading to a high-content eluate with no carryover of contaminants.