Kit
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Kit Content
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Catalog No.
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PAXgene Bone Marrow RNA Kit (30)
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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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764133
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Procedure
Bone marrow samples are collected using standard medical practices, with commonly used anticoagulants. A 2 ml sample is then transferred into a PAXgene Bone Marrow RNA Tube, which immediately stabilizes the RNA in the sample for storage at room temperature, refrigerated, or frozen. The samples can be safely transported in the PAXgene Bone Marrow RNA Tube, with no need for dry ice or liquid nitrogen. RNA can be purified days later using the PAXgene Bone Marrow RNA Kit* (see figure “RNA Stability in Bone Marrow Samples at Room Temperature”).
Principle
Copy numbers of individual mRNA species in bone marrow can change significantly during storage or transport at room temperature, making reliable studies of gene expression impossible. PAXgene Bone Marrow RNA Tubes protect RNA molecules from degradation by RNases and minimize ex vivo changes in gene expression (see figure “Stability of 5 Transcripts in Bone Marrow Samples at Room Temperature”), enabling accurate analysis of gene expression.
Rapid purification of high-quality RNA
The PAXgene Bone Marrow RNA Kit provides an easy and rapid procedure for isolation and purification of intracellular RNA from human whole bone marrow samples stabilized in PAXgene Bone Marrow RNA Tubes. The kit uses silica-membrane technology in a spin-column format for efficient purification of intact, high-quality RNA (see figures “RNA Stabilized During Storage at 2–8°C” and “High RNA Integrity after Freeze–Thaw Cycles”). An integrated DNase treatment ensures that genomic DNA is minimized. In-house and external studies showed 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 (see figure “RNA Purity for Samples Stored under Different Conditions”).
Efficient RNA purification from heterogeneous samples
Bone marrow samples are extremely heterogeneous, consisting of varying amounts of cells, tissue, and solid matter. RNA yields are highly donor-dependent and can vary greatly from sample to sample as well as between replicates from the same donor. The PAXgene Bone Marrow RNA Kit enables efficient isolation and purification of intracellular RNA from heterogeneous human whole bone marrow samples (see figure “RNA Yields from Bone Marrow Samples”), including an optimized procedure for cell-rich, viscous samples.
Applications
RNA purified using the PAXgene Bone Marrow RNA Kit is ready for use in a wide range of downstream research applications, including
• Real-time RT-PCR analysis of specific transcripts
• Array analysis of complex expression profiles
MSDS:
http://www1.qiagen.com/Support/Msds.aspx?catno=000000000000764114,000000000000764133
*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.
