PAXgene 96 Blood RNA Kit

• Parallel purification of RNA from 96 samples
• Standardized sample processing prior to analysis
• No organic extraction or precipitation
• Isolation of high-quality intracellular RNA
• Integrated DNase treatment for removal of gDNA

The PAXgene 96 Blood RNA Kit is for high-throughput purification of cellular RNA in a manual format. Used with PAXgene Blood RNA Tubes (available from BD, cat. no. 762165) for sample collection and stabilization, the system provides a complete preanalytical solution for collection and stabilization through to purification of high-quality RNA to support high-throughput projects, such as clinical research studies.

The PAXgene 96 Blood RNA Kit allows high-throughput isolation of total RNA from human whole blood. Together with PAXgene Blood RNA Tubes (available from BD, cat. no. 762165), it provides a complete preanalytical solution for collection and stabilization through to purification of high-quality RNA. The robust PAXgene 96 Blood RNA procedure is highly reproducible and typically yields between 4 and 20 µg of RNA from 2.5 ml human whole blood with minimal risk of sample-to-sample cross contamination. PAXgene Blood RNA Tubes contain a proprietary blend of reagents that bring about immediate stabilization of RNA. This blend prevents the drastic changes in the cellular RNA expression profiles that normally take place in vitro, after blood collection (see figures " RNA isolation from blood using standard methods", " Stabilization of gene expression", and " Detection of tyrosinase following blood storage"). The high-throughput procedure is simple, fast, and efficient, and processing 96 samples in parallel provides significant time savings in your clinical research studies.

In the PAXgene 96 Blood RNA System, blood is collected directly into PAXgene Blood RNA Tubes, and RNA is isolated using the PAXgene 96 Blood RNA Kit. PAXgene Blood RNA Tubes contain a proprietary blend of reagents that bring about immediate stabilization of RNA. Blood samples can be stored in the PAXgene Blood RNA Tubes for up to 5 days at room temperature. RNA is then isolated using silica membrane technology supplied in the PAXgene 96 Blood RNA Kit. The resulting RNA accurately represents the expression profile in vivo and is suitable for use in a range of downstream applications.

Blood samples (2.5 ml) are collected in PAXgene Blood RNA Tubes (available from BD, cat. no. 762165), and may be stored or transported at room temperature if desired. In the PAXgene 96 Blood RNA procedure, RNA isolation begins with a centrifugation step to pellet nucleic acids in the PAXgene Blood RNA Tube. The pellet is washed, and proteinase K is added to digest proteins. Alcohol is added to adjust binding conditions. Lysates are applied to the PAXgene 96 Filter Plate and centrifuged to remove cell debris. The lysate is then applied to a PAXgene 96 RNA Plate (see flowchart " PAXgene 96 Blood RNA procedure"). Residual DNA is removed through a DNase I digestion. Remaining contaminants are removed in three efficient wash steps before pure RNA is eluted.

The RNA isolated from human whole blood is ready for use in a wide range of downstream applications, including:

• Nucleic acid amplification technologies, such as RT-PCR and NASBA
• Expression-array and expression-chip analysis
• Quantitative RT-PCR, including TaqMan technology
• cDNA synthesis
• RNase and S1 nuclease protection
• Northern, dot, and slot blot analysis
• Primer extension