PAXgene Blood RNA MDx Kit (4)
For 4 x 96 RNA preps on the BioRobot MDx or Universal System workstation: 4 PAXgene 96 RNA Plates, 4 PAXgene 96 Filter Plates, Buffers (wash buffers are labeled with bar codes), Proteinase K, RNase-Free DNase Set, Plasticware, Collection Vessels. To be used with PAXgene Blood RNA Tubes
The PAXgene Blood RNA MDx Kit* is for fully automated, high-throughput purification of cellular RNA on the BioRobot® MDx or BioRobot Universal System. Used with PAXgene Blood RNA Tubes 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 trials or other large research studies.
Benefits of the PAXgene Blood RNA MDx System:
• Instant stabilization of RNA at point of collection — integrated system for use with PAXgene Blood RNA Tubes
• Optimized purification of high-quality RNA — standardized, automated processing of 48 or 96 samples in parallel
• Fully automated RNA purification procedure — on the BioRobot MDx or BioRobot Universal System
Integrated system for collection, stabilization, and automated high-throughput purification
Blood samples (2.5 ml) are collected in PAXgene Blood RNA Tubes, and may be stored or transported at room temperature if desired (Müller, M. C. et al  Leukemia 16, 2395; Rainen L. et al.  Clin Chem 48, 1883). The simple purification procedure begins with a centrifugation step to pellet the contents of each PAXgene Blood RNA Tube. Proteinase K and an optimized incubation buffer are added to digest proteins. The tubes are then transferred to the BioRobot MDx or BioRobot Universal System.
After setup of buffers and plasticware, guided by the QIAsoft Operating System, the BioRobot workstation carries out the fully automated RNA purification procedure (see figure “Automated RNA Purification”). Following purification, a final heat treatment of the eluate enhances performance in downstream applications. Total processing time is 3–4 hours, with a maximum of 25–30 minutes of hands-on time. Protocols are available for batches of 48 or 96 samples per run.
High-quality RNA with efficient DNase digestion
The automated PAXgene Blood RNA procedure provides high-quality RNA with high intra-donor repeatability (see figure “High-Quality RNA from Different Donors”). Since yields are highly donor-dependent, individual yields may vary. An integrated DNase treatment ensures that genomic DNA contamination is minimized, typically to <0.1% (see figure “Very Low Levels of Genomic DNA”). The robust and standardized procedure is designed to minimize the risk of sample-to-sample cross-contamination (see figure “No Detectable Cross-Contamination”).
High yields of RNA for all downstream applications
The fully automated PAXgene Blood RNA procedure provides RNA yields comparable to the manual PAXgene 96 Blood RNA procedure (typically ≥80%). Typical yields of RNA isolated from 2.5 ml healthy, human, whole blood (4.6 x 106 – 1.1 x 107 leukocytes/ml) are ≥3 μg for >95% of the samples processed. Purified RNA is ready to use in downstream applications, such as quantitative, real-time RT-PCR, with no RT-PCR inhibition detected (see figure “No RT-PCR Inhbition Detected”). Intersystem variation is minimal and well within the error range of typical real-time RT-PCR assays (see figure “High Intersystem Reproducibility”).
The purified RNA is ready for use in a wide range of downstream research applications, including:
• High-throughput, quantitative, real-time RT-PCR
• Expression-array and expression-chip analysis
• cDNA synthesis
• RNase and S1 nuclease protection
• Northern, dot, and slot blot analysis
• Primer extension
*The PAXgene Blood RNA MDx Kit is intended 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.