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PAXgene 96 Blood RNA Kit

For high-throughput purification of cellular RNA from whole blood
  • Parallel purification of RNA from 96 samples
  • Standardized sample processing prior to analysis
  • No organic extraction or precipitation
  • Isolation of high-quality cellular RNA

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.

Cat No./ID: 762331
PAXgene 96 Blood RNA Kit (4)
$4,091.00
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4 PAXgene 96 RNA Plates, 4 PAXgene 96 Filter Plates, Buffers, Proteinase K, RNase-free DNase Sets, AirPore Tape Sheets, Collection Vessels. To be used in conjunction with PAXgene Blood RNA Tubes.
Cat No./ID: 9238279
PAXgene 96 Incubator Block
$345.00
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Block for denaturation of eluates in PAXgene 96 procedures
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.

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RNA isolation from blood using standard methods.

Blood was collected and RNA was isolated using standard methods (collection in EDTA tubes; no stabilization; RNA isolation using a guanidinium-based method). The graph shows changes in expression of 12 genes after blood collection, measured using real-time RT-PCR. (Data kindly provided by Source Precision Medicine, Boulder, Colorado, USA.)

1

Stabilization of gene expression.

Blood was collected and RNA was isolated using the PAXgene Blood RNA System (for RNA stabilization and isolation). The graph shows changes in expression of 12 genes after blood collection, measured using real-time RT-PCR. (Data kindly provided by Source Precision Medicine, Boulder, Colorado, USA.)

2
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. Residual DNA is removed through a DNase I digestion. Remaining contaminants are removed in three efficient wash steps before pure RNA is eluted.
3
Detection of tyrosinase following blood storage.
Blood samples were collected from 6 different donors, spiked with tumor cells expressing tyrosinase, and stored for up to 10 days at room temperature. RT-PCR was used to measure levels of tyrosinase.  [A]  1 cell per 2.5 ml blood;  [B]  10 cells per 2.5 ml blood. WB: tyrosinase detection in RNA isolated from whole blood collected and stored in standard EDTA tubes. PAX: tyrosinase detection using the PAXgene Blood RNA System for sample collection, and RNA stabilization and isolation. (Data kindly provided by LMO, Munich, Germany.)
Performance
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.
Principle

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.

Procedure
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.
Applications

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
Features
Specifications
Applications PCR, real-time PCR, qPCR, NASBA, cDNA synthesis
CE/FDA/IVD compatible Research use only
Elution volume 45–60 µl
Format 96-well plate
Main sample type Whole blood
Processing Manual (centrifugation or vacuum)
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein Cellular RNA
Sample amount 2.5 ml
Stabilization Yes
Technology Silica technology
Time per run or per prep <1.5 hours
Yield 4–20 µg
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