For purification of DNA from tissues fixed and stabilized with the PAXgene Tissue System
Effective purification of high-quality, unmodified DNA
Preserved tissue morphology
Protocols for paraffin-embedded and paraffin-free tissues
Automated purification on the QIAcube
The PAXgene Tissue DNA Kit is intended for the purification of DNA 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 also includes the PAXgene Tissue FIX Container and PAXgene Tissue STABILIZER Concentrate for the collection, fixation, stabilization and storage of tissues samples.
For 50 DNA preps: PAXgene DNA Mini Spin Columns, Processing Tubes, Microcentrifuge Tubes, Carrier RNA, and Buffers; to be used in conjunction with PAXgene Tissue Containers
Specifications for fixation and storage conditions in PAXgene Tissue Fix and PAXgene Tissue Stabilizer were determined using animal tissues.
Significant hands-on time savings compared to the manual purification.
Comparison of sample preparation times between manual and QIAcube procedures for 12 PAXgene Tissue-fixed, paraffin-embedded (PFPE) tissues. Hands-on time without incubation times.
High-quality DNA from different tissue types fixed and stabilized in PAXgene Tissue Containers.
Tissues from different organs were fixed for 3 hours in PAXgene Tissue Fix, transferred to PAXgene Tissue Stabilizer, and stored for 24 hours at room temperature. After processing, DNA was purified from sections of paraffin-embedded tissue using the PAXgene Tissue DNA Kit. M: markers.
Yield of high-quality, high molecular weight DNA 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, DNA-rich and DNA-poor tissues, processed with the PAXgene Tissue DNA Kit on the QIAcube in one single run. DNA yield per 10 mg tissue.
Absorbance ratio of high-quality, high molecular weight DNA from 12 different PAXgene Tissue-fixed, paraffin-embedded (PF) tissue types, processed on the QIAcube.
Twelve different PAXgene Tissue-fixed (PF) rat tissue types, including fibrous or fatty, DNA-rich and DNA-poor tissue, processed with the PAXgene Tissue DNA Kit on the QIAcube in a single run. Ratio of absorbance at 260 and 280 nm.
Comparison of manual and automated procedure: DNA yield from sections of PAXgene Tissue-fixed, paraffin-embedded (PFPE) tissue.
DNA yield from 3 sections (thickness 10 µm) each of 8 different PAXgene Tissue-fixed, paraffin-embedded (PFPE) rat tissue types processed in triplicate with the PAXgene Tissue DNA Kit on the QIAcube or manually.
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 3 sections (thickness 10 µm) each of 8 different PAXgene Tissue-fixed, paraffin-embedded (PFPE) rat tissue types processed in triplicate with the PAXgene Tissue DNA Kit on the QIAcube or manually.
High-quality DNA from PFPE tissue with preserved morphology.
[A] Hematoxylin and eosin (H&E) staining of PAXgene Tissue-fixed, paraffin-embedded (PFPE) human colorectal cancer tissue and [B] DNA on agarose gel electrophoresis using 0.8% TBE buffered gels with 200 ng genomic DNA isolated in triplicate from 5 cases (#1–5) of human PFPE colorectral cancer. M: markers.
The PAXgene Tissue DNA procedure.
DNA without chemical modification can be used for demanding downstream applications.
Multiplex and long-range PCR of DNA from PAXgene Tissue-fixed, paraffin-embedded (PFPE) human colorectal cancer tissue (modified according to Viertler et al., (2012) A new technology for stabilization of biomolecules in tissues for combined histological and molecular analyses. J. MOl. Diagn. 14, 458). [A] Multiplex PCR of 8 different genomic DNA fragments ranging from 22 to 955 bp. [B] Long-range PCR of a 5 kb genomic DNA fragment.
Agarose gel electrophoresis of high-quality, high molecular weight DNA 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, DNA-rich and DNA-poor tissue, processed with the PAXgene Tissue DNA Kit on the QIAcube in one single run. DNA from each tissue type (300 ng) on agarose gel electrophoresis using 1% TBE buffered gels; M: markers.
Comparison of manual and automated procedure: Agarose gel electrophoresis from sections of PAXgene Tissue-fixed, paraffin-embedded (PFPE) tissue.
Agarose gel electrophoresis from 3 sections (thickness 10 µm) each of 8 different PAXgene Tissue-fixed, paraffin-embedded (PFPE) rat tissue types processed in triplicate with the PAXgene Tissue DNA Kit on the QIAcube and manually. DNA from each replicate and tissue type (200 ng) on agarose gel electrophoresis using 1% TBE buffered gels; M: markers.
The PAXgene Tissue DNA Kit enables purification of DNA from tissues fixed and stabilized using the PAXgene Fixation and Stabilization reagents, which preserve tissue morphology and biomolecule integrity by avoiding destructive crosslinking and degradation found in formalin-fixed tissues. The purified DNA 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 lysed in Buffer TD1 with proteinase K. Conditions are adjusted for optimal binding of DNA to the silica membrane and the lysate is loaded on the PAXgene DNA spin column. DNA binds to the membrane while contaminants pass through. Enzyme inhibitors are effectively removed with subsequent washes and pure DNA is eluted in a low-salt elution buffer (see The PAXgene Tissue DNA procedure).
The PAXgene Tissue DNA Kit provides 2 protocols for purification of DNA from different starting materials: from sections of PFPE tissues and from PF tissue samples (without paraffin embedding).
Automation
DNA purification using the PAXgene Tissue DNA Kit can be uatomated on the QIAcube, enabling:
Two protocols are available for automated purification from different starting materials:
DNA from PFPE tissue sections, staring with pellets of 2–5 deparaffinized PFPE tissue sections
DNA from tissue samples fixed and stabilized in a PAXgene Tissue FIX Container, starting with lysates from up to 10 mg fixed and stabilized tissue, homogenized in lysis buffer TD1
Applications
The purified DNA is ready to use in a wide range of downstream applications, including:
PCR, multiplex, long-range and quantitative, real-time PCR
Southern blotting
SNP genotyping
Next-generation sequencing (NGS)
Features
Specifications
Applications
PCR and quantitative, real-time PCR, Southern blotting, pharmacogenomic studies, SNP discovery and SNP genotyping
Elution volume
14–40 µl
Format
Spin column
Main sample type
Human tissue
Processing
Manual (centrifugation)
Sample amount
4 x 15 x 15 mm
Technology
Silica technology
Time per run
30 min + 120 min incubation/8 samples
Yield
Depends on tissue type and starting material (fixed or PFPE*)
* PAXgene Fixed Paraffin Embedded.
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
