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PAXgene Tissue RNA Kit

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

  • Effective purification of high molecular weight RNA from tissue with preserved morphology
  • Protocols for paraffin-embedded or paraffin-free tissues
  • Minimal genomic DNA contamination
  • Automated purification on the QIAcube

 

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 of PFPE** tissue with a maximum size of 15 x 15 mm
Elution volume 14–40 µl
Time per run 60 min + 15 min incubation/8 samples
(120 min for fibrous tissues)
Processing Manual: centrifugation
Automated: QIAcube

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

 

Intended Use

The PAXgene Tissue RNA Kit is intended for the purification of RNA 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, and stabilization 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 RNA Procedure
Disruption and homogenization of the tissue sample is performed in binding buffer, Buffer TR1. After a centrifugation step to remove residual cell debris, ethanol is added to the lysate to provide [...]
High-quality RNA from PFPE tissue with preserved morphology
(A) Hematoxylin and eosin (H&E) staining of PAXgene Tissue fixed, paraffin-embedded (PFPE) rat tissue of different organs and (B) RNA from sections of rat PFPE. Electropherogram and RNA integrity [...]
RNA from PFPE tissue without chemical modifications
RNA from PFPE tissue without chemical modifications
SYBR Green real-time RT-qPCR with 10 ng RNA from cryo preserved, formalin-fixed, paraffin-embedded (FFPE), or rat PFPE tissue (modified according to Groelz et al. Exp Mol Pathol. 2013). Average [...]
Significant hands-on time savings compared to the manual purification.
Comparison of sample preparation times between manual and QIAcube procedures for preparation of 12 PAXgene Tissue fixed, paraffin-embedded (PFPE) samples. Hands-on time without incubation
RNA yield of high-quality, high molecular weight RNA from 12 different PAXgene Tissue fixed (PF) tissue types, processed on the QIAcube.
Twelve different PAXgene Tissue fixed (PF) rat tissue types, including fibrous or fatty, RNA rich and RNA poor tissue, processed with the PAXgene Tissue RNA kit on the QIAcube. RNA yield per 10 mg [...]
Absorbance ratio of high-quality, high molecular weight RNA from 12 different PAXgene Tissue fixed (PF) tissue types, processed on the QIAcube.
Twelve different PAXgene Tissue fixed (PF) rat tissue types, including fibrous or fatty, RNA rich and RNA poor tissue, processed with the PAXgene Tissue RNA kit on the QIAcube in one single run. [...]
RIN value of high-quality, high molecular weight RNA from 12 different PAXgene Tissue fixed (PF) tissue types, processed on the QIAcube.
Twelve different PAXgene Tissue fixed (PF) rat tissue types, including fibrous or fatty, RNA rich and RNA poor tissue, processed with the PAXgene Tissue RNA kit on the QIAcube in one single run. RNA [...]
Comparison of manual and automated procedure: RNA yield from sections of PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue.
RNA yield from 5 sections (thickness 10 µm) each of eight different PAXgene Tissue fixed, paraffin-embedded (PFPE) rat tissue types processed in triplicates with the PAXgene Tissue RNA kit on the [...]
Comparison of manual and automated procedure: Absorbance ratio from sections of PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue.
Ratio of absorbance at 260 and 280 nm from 5 sections (thickness 10 µm) each of eight different PAXgene Tissue fixed, paraffin-embedded (PFPE) rat tissue types processed in triplicates with the [...]
Comparison of manual and automated procedure: RIN value from sections of PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue.
Twelve different PAXgene Tissue fixed (PF) rat tissue types, including fibrous or fatty, RNA rich and RNA poor tissue, processed with the PAXgene Tissue RNA kit on the QIAcube in one single run. RNA [...]
Comparison of manual and automated procedure: Cycle treshold values from a reverse transcription and amplification from sections of PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue.
Twelve different PAXgene Tissue fixed (PF) rat tissue types, including fibrous or fatty, RNA rich and RNA poor tissue, processed with the PAXgene Tissue RNA kit on the QIAcube in one single run. [...]
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Performance

Total RNA purified using the PAXgene Tissue RNA Kit is highly pure. Genomic DNA contamination is minimized, and purified RNA is ready to use in downstream applications with no detectable inhibition of PCR (see High-quality RNA from PFPE tissue with preserved morphology and RNA from PFPE tissue without chemical modification). All RNA molecules longer than 200 nucleotides are purified. The procedure provides enrichment for mRNA since most RNAs <200 nucleotides (such as 5.8S rRNA, 5S rRNA, and tRNAs, which together comprise 15–20% of total RNA) are selectively excluded. The size distribution of the purified RNA is comparable to that obtained by centrifugation on a CsCl gradient, through which small RNAs do not sediment efficiently. The PAXgene Tissue miRNA Kit is recommended for purification of small RNAs, including miRNA.

 

Principle

The PAXgene Tissue RNA Kit enables purification of 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 has 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 to the silica membrane. Contaminants are then washed away and DNase I treatment removes any trace amounts of bound DNA. RNA is then eluted in a low-salt elution buffer and denatured by heating (see The PAXgene Tissue RNA Procedure).

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

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

 

Automation

RNA purification using the PAXgene Tissue RNA Kit can be automated on the QIAcube, enabling:

 

Two protocols are available for automated purification from different starting materials:

 

Applications

The purified RNA is ready to use in a wide range of downstream applications (see Resources), including:

  • cDNA synthesis
  • Gene expression arrays
  • End-point RT-PCR
  • Quantitative RT-PCR
  • RNA sequencing
 

Handbooks (1)

File sizeFormatDownload
PAXgene Tissue RNA Kit Handbook 372 KB pdf

Supplementary Protocols (4)

File sizeFormatDownload
Simultaneous purification of genomic DNA and total RNA from sections of PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue 84 KB pdf
Deparaffinization of PAXgene Tissue fixed, paraffin-embedded tissue sections with Deparaffinization Solution 86 KB pdf
Purification of total RNA from sections of PAXgene® Tissue fixed, cryo-embedded (PFCE) tissue placed directly into a microcentrifuge tube 603 KB pdf
Purification of total RNA from microdissected PAXgene® Tissue fixed, paraffin-embedded (PFPE) and PAXgene Tissue fixed, cryo-embedded (PFCE) tissues 735 KB pdf

Brochures (2)

File sizeFormatDownload
RNA Isolation with the PAXgene Tissue RNA Kit 2417 KB pdf
PAXgene Tissue System Brochure 4792 KB pdf

Technical information (7)

File sizeFormatDownload
Yield, purity, and integrity of RNA purified from PAXgene Tissue fixed, paraffin embedded (PFPE) rat tissue 618 KB pdf
Vaccum sealing of fixed and stabilized tissue with a FoodSaver Vacuum Sealer for dry and safe transport 886 KB pdf
Simultaneous preservation of RNA and morphology in tissue samples fixed with PAXgene Tissue Fix and stored for up to 2 years in PAXgene Tissue Stabilizer at –20 or –80°C 1313 KB pdf
RNA stability in tissue samples, fixed and stabilized with the PAXgene Tissue System, for up to 7 days at 22°C or 2 months at 4°C 538 KB pdf
Morphology and RNA preservation in PAXgene Tissue fixed, paraffin-embedded tissue (PFPE) stored for 18 months at different temperatures 495 KB pdf
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

Posters (14)

File sizeFormatDownload
New Fixation Technology For Simultaneous Preservation of Morphology and Nucleic Acids in Tissue; Groelz et al., AMP 2008 423 KB pdf
Long-Term Storage of Tissue Specimen at –20°C to –80°C with Preservation of Morphology and Nucleic Acids within Frozen Tissue; Groelz et al., AMP 2009/2 693 KB pdf
Preservation of Histomorphology and Nucleic Acids in Human Breast Tumor Tissue with the New PAXgene Tissue System — A Study with Comparison to Formalin Fixation; Groelz et al., AMP 2009/1 912 KB pdf
Preservation of Gene Expression Profile and Histomorphology in Human Breast Tumor Tissue with the New PAXgene Tissue System; Groelz et al., AACR 2010 769 KB pdf
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 vs. Formalin Fixed Tissue: A Comparison of Tissue Morphology and RNA Quality; Groelz el al., BRN Symposium 2012 5200 KB pdf
Preservation of Morphology and Biomolecules Within Tissue Stored for Three Years at -80°C in PAXgene Tissue Stabilizer Reagent; Groelz et al., ISBER 2012 912 KB pdf
PAXgene Tissue Fixation Technology for Simultaneous Preservation of Morphology and Biomolecules; Groelz et al., ECP 2012 1231 KB pdf
RNA and Morphology Preservation after 5 Years storage at -20 and 3 years at -80°C; Groelz et al., GTEx Meeting 2014 909 KB pdf
Stability of Nucleic Acids in Archived Formalin and PAXgene Tissue Fixed, Paraffin-embedded Blocks of Tissue; Groelz et al., ECP 2014 2617 KB pdf
RT-PCR Performance of RNA Obtained From Archived FFPE and PFPE Blocks of Tissue; Groelz et al., AMP 2014 7568 KB pdf
RT-PCR Performance of RNA Obtained From Archived FFPE and PFPE Blocks of Tissue; Groelz et al., GTEx Meeting 2015 7568 KB pdf

References (9)

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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. Link
The Genotype-Tissue Expression (GTEx) Project: Linking Clinical Data with Molecular Analysis to Advance Personalized Medicine; Keen et al., J. Pers. Med. 2015, 5, 22-29 Link
ProductCatalog Nr.Price (USD)

PAXgene Tissue RNA 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.
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