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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
  • Integrated DNase treatment for removal of gDNA
  • No organic extraction or precipitation
  • Isolation of high-quality intracellular RNA
FeatureSpecification
Format

96-well plate

Technology

Silica membrane

Sample amount 

2.5 ml

Elution volume

90–120 µl

Main sample type

Whole blood

Time per run

<3.5 h

Typical RNA yield

4–20 µg/2.5 ml sample

RNA purity  

1.8–2.2 (A260/A280)

Processing 

Manual: centrifugation or vacuum


Intended Use

The PAXgene 96 Blood RNA Kit is intended for the purification of cellular RNA from whole blood collected in PAXgene Blood RNA Tubes.

For research use only. Not for use in diagnostic procedures. No claim or representation is intended to provide information for the diagnosis, prevention, or treatment of a disease. The performance characteristics of this product have not been fully established.

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PAXgene 96 Blood RNA Kit

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Details

Performance

The PAXgene 96 Blood RNA Kit allows high-throughput isolation of total RNA from human whole blood. The robust PAXgene 96 Blood RNA procedure is highly reproducible and typically yields are between 4 and 20 µg of RNA from 2.5 ml human whole blood with minimal risk of sample-to-sample cross contamination. The high-throughput procedure is simple, fast, and efficient, and processing 96 samples in parallel provides significant time savings.

Principle

Copy numbers of individual RNA species in blood can change significantly during storage or transport at ambient temperature, making reliable studies of gene expression impossible. Both stabilization of RNA molecules after collection and an efficient RNA purification protocol are needed to maximize insights into expression profiles of whole blood. Based on proven silica-membrane technology in a high-throughput, 96-well plate format, and when combined with PAXgene Blood RNA Tubes, the PAXgene 96 Blood RNA Kit provides the chemistry and rapid procedure to isolate RNA of high quality and purity.

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 (Figure 1). 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 purified RNA is ready for use in a wide range of downstream applications, including:

  • cDNA synthesis
  • Gene expression arrays
  • End-point RT-PCR
  • Quantitative RT-PCR, including TaqMan technology
  • Next-generation sequencing
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.Zoom image

Figure 1. PAXgene 96 Blood RNA procedure.

RNA isolation begins with a centrifugation step to pellet nucleic acids in the ...

Resources

PAXgene 96 Blood RNA Kit


1. Can the PAXgene 96 Blood RNA Kit be used for diagnostic or prognostic procedures?
No. The product is ‘For Research Use.’ Not for use in diagnostic procedures. No claim or representation is intended to provide information for the diagnosis, prevention, or treatment of a disease?

2. How many PAXgene Blood RNA Tubes can be processed per PAXgene RNA 96 well plate?
The PAXgene RNA 96 well plate purification chemistry was designed to link to the PAXgene Blood RNA Tubes, therefore, one PAXgene Blood RNA Tube can be processed per well or 96 tubes per plate.

3. What is the PAXgene 96 filter plate?
The PAXgene 96 filter plate is a special device that filters contaminants and generates a cleared lysate during centrifugation.

4. What additional equipment is needed to purify RNA using the PAXgene 96 Blood RNA Kit?
The PAXgene 96 Blood RNA Kit uses a combination of vacuum and centrifugation to isolate RNA. The vacuum manifold required is the QIAGEN QIAvac 96 vacuum manifold, cat. no. 19504. The suggested centrifuge is manufactured by Sigma and sold by QIAGEN (Centrifuge 4-16KS; cat. nos. 81600, 81610, 81620 and 81625), as is the additional plate rotor for spinning 2 x 96 plates, cat. no. 81031. Additionally, the final heating step of the protocol utilizes an alloy block, the PAXgene 96 Incubation block, cat. no. 9238279. Please contact QIAGEN technical service (www.qiagen.com) for more information.

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