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PAXgene Tissue miRNA Kit

For isolation and purification of total RNA, including miRNA, from tissues fixed and stabilized using the PAXgene Tissue Container

  • Effective purification of high and low molecular weight RNA from tissue with preserved morphology
  • Protocols for paraffin-embedded or paraffin-free tissues
  • Minimal genomic DNA contamination
  • Reliable quantification of miRNAs

 

Feature Specification
Format Spin column
Technology Silica membrane
Main sample type Human tissue
Sample amount Up to 10 mg from PF* tissue
2–5 sections 10 µm thick from PFPE** tissue with a maximum
size of 15 x 15 mm
Elution volume 14–40 µl
Time per run 70 min + 15 min incubation/8 samples
(120 min for fibrous tissues)
Processing Manual: centrifugation

* PAXgene Tissue fixed
** PAXgene Tissue fixed, paraffin-embedded

 

Intended Use

The PAXgene Tissue miRNA Kit is intended for the purification of total RNA, including miRNA, from PAXgene Tissue fixed (PF) and PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue samples. The kit is intended to be used as part of the PAXgene Tissue System, which additionally includes the PAXgene Tissue Container or PAXgene Tissue FIX Container and the PAXgene Tissue STABILIZER Concentrate for the collection, fixation, stabilization, and storage of tissue samples.

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.

The PAXgene Tissue miRNA Procedure
Disruption and homogenization of the tissue sample is performed in binding buffer, Buffer TM1. After centrifugation to remove residual cell debris, isopropanol is added to the lysate to provide [...]
Efficient purification of miRNA from PFPE tissue with the PAXgene Tissue miRNA Kit
RNA purification from sections of six different rat PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue types using the PAXgene Tissue RNA Kit or PAXgene Tissue miRNA Kit. Amplification in a [...]
High concordance of miRNA expression between total RNA isolated from PFPE and flash-frozen tissue
miRNA purified from mirrored human breast cancer tissue fresh frozen in liquid nitrogen using the QIAGEN miRNeasy Kit, or from sections of PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue using [...]
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Performance

Total RNA purifed using the PAXgene Tissue miRNA Kit is highly pure. Genomic DNA contamination is minimized and the purified RNA is ready to use in downstream applications with no detectable inhibition of PCR. The resulting eluate includes smaller RNA molecules, such as 5.8S rRNA, 5S rRNA, tRNAs, and miRNAs. Studies have shown that miRNA purified from PFPE tissue samples is reliably quantified, with high concordance to flash-frozen samples (see Efficient purification of miRNA from PFPE tissue with the PAXgene Tissue miRNA Kit and High concordance of miRNA expression between total RNA isolated from PFPE and flash-frozen tissue).

 

Principle

The PAXgene Tissue miRNA Kit enables purification of total RNA from tissues fixed and stabilized using the PAXgene Fixation and Stabilization products, which preserve tissue morphology and biomolecule integrity by avoiding destructive cross-linking and degradation found in formalin-fixed tissues. The purified RNA and miRNA have no inhibitory chemical modifications and thus, can be used in sensitive downstream applications.

 

Procedure

PAxgene Tissue fixed (PF) or PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue samples are disrupted and homogenized in binding buffer. After centrifugation to remove cellular debris, optimal conditions are created for binding of RNA molecules to the silica membrane. Contaminants are then washed away and DNase I treatment removes any trace amounts of bound DNA. Total RNA, including miRNA, is then eluted in a low-salt elution buffer and denatured by heating (see The PAXgene Tissue miRNA Procedure).

 

The PAXgene Tissue miRNA Kit provides 3 protocols for purification of RNA, including miRNA, from different starting materials:

  • Sections of PFPE tissues
  • PF tissue samples (without paraffin embedding)
  • Blocks of PFPE tissue samples

 

Applications

Total RNA and miRNA purified with the PAXgene Tissue miRNA Kit is ready to use in a range of downstream research applications (see Resources), including:

  • cDNA synthesis
  • Gene expression arrays
  • End-point RT-PCR
  • Quantitative RT-PCR
  • Detection and quantification of miRNA
  • RNA sequencing
 

Handbooks (1)

File sizeFormatDownload
PAXgene Tissue miRNA Handbook 381 KB pdf

Supplementary Protocols (4)

File sizeFormatDownload
Purification of total RNA, including miRNA, from sections of PAXgene Tissue fixed, cryo-embedded (PFCE) tissue placed directly into a microcentrifuge tube 621 KB pdf
Simultaneous purification of genomic DNA and total RNA, including miRNA, from sections of PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue 85 KB pdf
Deparaffinization of PAXgene Tissue fixed, paraffin-embedded tissue sections with Deparaffinization Solution 86 KB pdf
Purification of total RNA, including miRNA, from microdissected PAXgene® Tissue fixed, paraffin embedded (PFPE) and PAXgene Tissue fixed, cryoembedded (PFCE) tissues 679 KB pdf

Brochures (2)

File sizeFormatDownload
miRNA Isolation with the PAXgene Tissue miRNA Kit 957 KB pdf
PAXgene Tissue System Brochure 4792 KB pdf

Technical information (2)

File sizeFormatDownload
Influence on RNA yield and integrity of modifications to the processing protocol 648 KB pdf
Influence of formalin contamination during processing of PAXgene Tissue fixed, paraffin-embedded tissue (PFPE) on RNA yield, integrity, and performance in quantitative RT-PCR 677 KB pdf

Safety Data Sheets (1)

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MSDS PAXgene Tissue miRNA Kit Link

Posters (4)

File sizeFormatDownload
A New Tissue Fixative for Biomarker Discovery: Gene expression and miRNA in PAXgene® Tissue Fixed Paraffin-Embedded (PFPE) Colorectal Cancer (CRC) and Breast Cancer Tissue vs. Formalin Fixed Paraffin-Embedded (FFPE) Tissue; Groelz et al., 3rd Annual Oncology Biomarkers 2011 423 KB pdf
PAXgene Tissue: A New Fixation Technology for Simultaneous Preservation of Morphology and Nucleic Acids; Groelz et al., BRN Symposium 2011 555 KB pdf
Evaluation of the PAXgene® Tissue System: Preservation of morphology and gene expression in human melanoma; Hesse et al., AACR-NCI-EORTC 2011 997 KB pdf
PAXgene Tissue Fixation Technology for Simultaneous Preservation of Morphology and Biomolecules; Groelz et al., ECP 2012 1231 KB pdf

References (9)

File sizeFormatDownload
Feature-based analysis of mouse prostatic intraepithelial neoplasia in histological tissue sections. Ruusuvuori P. et al., J Pathol Inform. 2016 Jan 29;7:5 Link
Pathogen Inactivating Properties and Increased Sensitivity in Molecular Diagnostics by PAXgene, a Novel Non-Crosslinking Tissue Fixative. Loibner M. et al., PLoS One. 2016 Mar 14;11(3):e0151383. Link
Protocol for HER2 FISH determination on PAXgene-fixed and paraffin-embedded tissue in breast cancer. Oberauner-Wappis L et al., Int J Exp Pathol. 2016 Apr;97(2):202-6 Link
The focus on sample quality: Influence of colon tissue collection on reliability of qPCR data. Korenkova V et al., Sci Rep. 2016 Jul 7;6:29023. doi: 10.1038/srep29023 Link
A Critical Evaluation of the PAXgene Tissue Fixation System:  Morphology, Immunohistochemistry, Molecular Biology, and Proteomics. Mathieson W. et al., Am J Clin Pathol. 2016 Jul;146(1):25-40. Link
Assessment of PAXgene Fixation on Preservation of Morphology and Nucleic Acids in Microdissected Retina Tissue. Liu Y and Edward DP, Curr Eye Res. 2016 Jul 13:1-7. Link
Surgical Specimens of Colorectal Cancer Fixed with PAXgene Tissue System Preserve High-Quality RNA; Hara K. et al., Biopreserv Biobank. 2015 Oct;13(5):325-34 Link
A Novel Approach to High-Quality Postmortem Tissue Procurement: The GTEx Project; Carithers et al., Biopreservation and Biobanking. October 2015, 13(5): 311-319 931 KB pdf
Next-gen tissue: preservation of molecular and morphological fidelity in prostate tissue; Gillard M. et al., Am J Transl Res 2015;7(7):1227-1235 Link
ProductCatalog Nr.Price (USD)

PAXgene Tissue miRNA Kit (50)

For 50 RNA preps: PAXgene RNA MinElute Spin Columns, PAXgene Shredder Spin Columns, Processing Tubes, Microcentrifuge Tubes, Carrier RNA, RNase-Free DNase, and RNase-Free Buffers. To be used in conjunction with PAXgene Tissue Containers or PAXgene Tissue FIX Containers + PAXgene Tissue STABILIZER reagent.
766134610.00