text.skipToContent text.skipToNavigation

PAXgene Tissue miRNA Kit

For purification of microRNA and total RNA from tissues fixed and stabilized in PAXgene Tissue Containers
  • Integrated fixation, stabilization, and purification
  • Effective purification of miRNA and total RNA
  • High-quality miRNA from tissues
  • Preserved tissue morphology

Tissue samples fixed and stored in PAXgene Tissue Containers can be paraffin-embedded and used for pathological studies as well as for subsequent purification of miRNA, RNA, and/or DNA. The PAXgene Tissue miRNA Kit provides purification of total RNA, including RNA from approximately 18 nucleotides, from tissues fixed and stabilized in PAXgene Tissue Containers. Purification is carried out using silica-based RNA purification technology in a spin-column format. Used with the containers, the kit provides a complete preanalytical solution for collection, fixation, and stabilization through to purification of high-quality miRNA and total RNA for molecular analysis.

Cat No./ID: 766134
PAXgene Tissue miRNA Kit (50)
$972.00
Add To Cart
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
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.
0
Efficient purification of miRNA from fixed tissue stored 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, total RNA was purified from sections of paraffin-embedded tissue using the PAXgene Tissue RNA Kit (RNA/miRNA). miRNA-enriched fractions (200 nt) were also isolated from sections of the same tissue blocks using the PAXgene Tissue miRNA Kit (Enriched miRNA). Purified RNA was used as a template in quantitative, real-time RT-PCR assays for the miRNA miR-10a using the miScript Reverse Transcription Kit and the miScript SYBR® Green PCR Kit.
1
The PAXgene Tissue miRNA procedure.
Performance

Together the container and kit provide a complete preanalytical solution for collection, fixation, and stabilization of tissue, and for purification of high-quality RNA, including miRNA, for molecular analysis (see figure "Efficient purification of miRNA from fixed tissue stored in PAXgene Tissue Containers").

Total RNA purified using the PAXgene Tissue miRNA Kit is highly pure. Genomic DNA contamination is minimized, and purified RNA is ready to use in downstream applications with no detectable PCR inhibition. All RNA molecules longer than 18 nucleotides are purified.

Principle

Current tissue fixation methods used in traditional histology are of limited use for molecular analysis. Fixatives that contain formaldehyde cross-link biomolecules and modify nucleic acids and proteins. During tissue fixation, storage, and processing, cross-links lead to degradation of nucleic acids. Since cross-links can not be removed completely, the resulting chemical modifications can cause inhibition in sensitive downstream applications such as quantitative PCR or RT-PCR. In order to enable both molecular and traditional pathology testing from the same specimen, a method is needed for stabilization of molecular content and preservation of morphology.

PreAnalytiX has developed the PAXgene Tissue System to meet such needs. The system consists of a tissue collection device (the PAXgene Tissue Container for collection, stabilization, storage, and transportation of human tissue specimens) and kits for purification of miRNA, total RNA, or DNA. PAXgene Tissue Containers provide tissue fixation for histopathology studies and enable purification of high-quality nucleic acids from the same sample for molecular analysis. The fixation and stabilization method preserves tissue morphology and the integrity of nucleic acids without destructive cross-linking and degradation found in formalin-fixed tissues.

For purification of total RNA including miRNA, the system requires the use of PAXgene Tissue Containers for tissue collection and stabilization, followed by RNA isolation and purification using the PAXgene Tissue miRNA Kit.
Procedure

PAXgene Tissue Containers are dual-chamber containers prefilled with 2 reagents. PAXgene Tissue Fix rapidly penetrates and fixes the tissue. After fixation, the tissue is removed from the PAXgene Tissue Fix and transferred to PAXgene Tissue Stabilizer in the second chamber of the same container. When the tissue is stored in PAXgene Tissue Stabilizer, nucleic acids and morphology of the tissue sample are stable for a minimum of 3 and a maximum of 7 days at room temperature or for up to 8 weeks at 2–8°C, depending on the type of tissue. Storage at –15 to –30°C is also possible for at least 26 months without any negative effects on the morphology of the tissue or the integrity of the nucleic acids.

Stabilized samples can be embedded in paraffin for histological studies. Nucleic acids can be isolated from the PAXgene Tissue fixed, paraffin-embedded (PFPE) samples either before or after embedding in paraffin.

The PAXgene Tissue miRNA Kit provides 3 protocols for purification of total RNA from tissue fixed and stabilized in PAXgene Tissue Containers, including RNA molecules smaller than 200 nucleotides, such as 5.8S rRNA, 5S rRNA, tRNAs, and miRNAs. Optimized binding and washing conditions ensure the purification of RNA molecules as small as 18 nucleotides. As a prerequisite, the tissue must be fixed and stabilized in PAXgene Tissue Containers.

Disruption and homogenization of the tissue sample is performed in the binding buffer, Buffer TM1. After a centrifugation step to remove residual cell debris, ethanol is added to the lysate to provide appropriate binding conditions for all RNA molecules 18 nucleotides and longer. The sample is then applied to a PAXgene RNA MinElute spin column where the total RNA binds to the membrane and contaminants are efficiently washed away. Between the first and second wash steps, the membrane is treated with DNase I to remove trace amounts of bound DNA. After the wash steps, RNA including miRNA is eluted in a low-salt elution buffer and denatured by heating (see figure "The PAXgene Tissue miRNA procedure").

Specifications for fixation and storage conditions in PAXgene Tissue Fix and PAXgene Tissue Stabilizer were determined using animal tissues.

Applications

The purified miRNA and total RNA is ready to use in a wide range of downstream applications, including:

  • Northern blot analysis
  • RT-PCR and quantitative, real-time RT-PCR
  • Microarray analysis
Features
Specifications
Applications null
Applications null
Elution volume null
Elution volume null
Format null
Format null
Main sample type null
Main sample type null
Processing null
Processing null
Sample amount null
Sample amount null
Technology null
Technology null
Time per run null
Time per run null
Yield null
Yield null

You are not authorized to download the resource

Brochures & Guides (4)
Two worlds in one sample
Show details

Show details
For isolation and purification of total RNA, including miRNA, from tissue samples fixed and stabilized using the PAXgene Tissue System

 


Show details
Moving toward excellence and standardization in tissue collection and fixation

 


Show details
Kit Handbooks (1)
For isolation and purification of total RNA, including miRNA, from tissue samples stabilized in PAXgene Tissue Containers
Show details
References
0
fragment fix placeholder

Customers who bought these products also bought

  • Cat No./ID: 217504
    /qiagenstorefront/au//sample-technologies/rna/mirneasy-ffpe-kit/

    miRNeasy FFPE Kit (50)

    50 RNeasy MinElute Spin Columns, Collection Tubes, Proteinase K, RNase-Free DNase I, DNase Booster Buffer, RNase-Free Buffers, RNase-Free Water
    $926.00
  • Cat No./ID: 767134
    /qiagenstorefront/au//sample-technologies/dna/paxgene-tissue-dna-kit/

    PAXgene Tissue DNA Kit (50)

    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
    $439.00