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PAXgene Bone Marrow RNA Kit

For purification of intracellular RNA from bone marrow samples in PAXgene Bone Marrow RNA Tubes

  • To be used in conjunction with the PAXgene Bone Marrow RNA Tube
  • Rapid purification of RNA with high integrity and purity
  • Optimized protocol for heterogeneous bone marrow samples
  • Integrated DNase treatment for gDNA removal

Feature

Specification

Format

Spin column

Technology

Silica membrane

Sample amount

2 ml

Elution volume

80 µl (2 x 40 µl)

Main sample type

Bone marrow aspirates

Time per run

95 min/12 samples

Median RNA yield

11 µg/2 ml sample (n=295)

RNA purity

1.8–2.2 (A260/A280)

Processing

Manual: centrifugation


Intended Use

The PAXgene Bone Marrow RNA System consists of a bone marrow collection tube (PAXgene Bone Marrow RNA Tube) and nucleic acid preparation kit (PAXgene Bone Marrow RNA Kit). It is intended for the collection, storage and transport of human bone marrow, stabilization of intracellular RNA in a closed tube and subsequent isolation and purification of total RNA.

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.

Order information

Product

Catalog Nr.

Price

PAXgene Bone Marrow RNA Kit (30)

For 30 RNA preps: 30 PAXgene Spin Columns, 30 PAXgene Shredder Spin Columns, Processing Tubes, RNase-free DNase I, RNase-free reagents and buffers. To be used in conjunction with PAXgene Bone Marrow RNA Tubes.

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Details

Performance

The PAXgene Bone Marrow RNA Kit enables efficient isolation and purification of intracellular RNA from heterogeneous human whole bone marrow samples that have been stabilized in the PAXgene Bone Marrow RNA Tube (Figure 1). The process includes an optimized procedure for cell-rich, viscous samples. RNA can be purified days after sample stabilization in PAXgene Bone Marrow RNA Tubes. An integrated DNase treatment step ensures that genomic DNA is depleted from samples. In-house and external studies have shown that genomic DNA was <0.85% in 95% of the samples tested (n = 157). A260/A280 values are typically between 1.8 and 2.2, indicating the high purity of the RNA. Combining the PAXgene Bone Marrow RNA Kit with PAXgene Bone Marrow RNA Tubes ensures that RNA is protected from degradation by RNases. Furthermore, stabilization enables accurate gene expression analysis because ex vivo changes in transcript levels due to gene regulation and RNA degradation are minimized (Figure 2).

Principle

Copy numbers of individual mRNA species in bone marrow 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 bone marrow aspirates. Based on silica-membrane technology in a spin-column format, the PAXgene Bone Marrow RNA Kit provides the chemistry and rapid procedure to isolate RNA of high quality and purity from bone marrow aspirate samples stabilized in PAXgene Bone Marrow RNA Tubes.

Procedure

A bone marrow aspirate sample (2 ml) is introduced into the PAXgene Bone Marrow RNA Tube and incubated at room temperature for a minimum of 2 hours or up to 3 days. Nucleic acids are concentrated by centrifugation and the resuspended precipitate is treated with proteinase K. RNA is then allowed to bind to the silica membrane and contaminants are removed by washing. A DNase treatment step subsequently removes DNA, leaving pure RNA for final elution (Figure 3).

Applications

RNA purified using the PAXgene Bone Marrow RNA Kit 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

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