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miRNeasy FFPE Kit (50)
Cat no. / ID. 217504
50 RNeasy MinElute Spin Columns, Collection Tubes, Proteinase K, RNase-Free DNase I, DNase Booster Buffer, RNase-Free Buffers, RNase-Free Water
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The miRNeasy FFPE Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
Want to try this solution for the first time?
Get in touch with our team today and request a quote to trial the miRNeasy FFPE Kit (50).
Features
- Effective purification of miRNA and total RNA
- Novel method to overcome formalin crosslinking
- Efficient release of RNA without compromising integrity
- Streamlined protocol providing RNA in just 85 minutes
Product Details
The miRNeasy FFPE Kit enables purification of total RNA that includes RNA from approximately 18 nucleotides upwards from formalin-fixed, paraffin-embedded (FFPE) tissue sections. The kit provides recovery of usable RNA fragments, including miRNA and other small RNAs, for applications such as quantitative, real-time RT-PCR. Procedures can be automated on the QIAcube Connect.
Performance
Yields and performance of total RNA and miRNA with the miRNeasy FFPE Kit are superior to alternative methods of miRNA purification, such as using phenol-chloroform extraction or a similar kit from another supplier (see figures " Effective real-time RT-PCR quantification", " Efficient RNA purification from FFPE tissues" and Superior yields and performance).
See figures
Principle
Fixing tissues with formalin leads to RNA-RNA and RNA-protein crosslinking which impairs RNA performance in downstream applications. The miRNeasy FFPE Kit provides special lysis and incubation conditions to reverse formalin crosslinking of RNA. A specially developed lysis buffer efficiently releases RNA from tissue sections while avoiding further RNA degradation. The lysate is treated with DNase then optimized binding conditions allow purification of all usable RNA from approximately 18 nucleotides upwards.
Procedure
The entire miRNeasy FFPE procedure can be completed in as little as 85 minutes. The optimized lysis buffer allows sample lysis with proteinase K digestion in only 15 minutes without sacrificing RNA yields. After lysis, samples are incubated at 80ºC for 15 minutes to reverse formalin crosslinking. Genomic DNA is then rapidly removed in an optimized DNase step with DNase Booster Buffer, and concentrated RNA is purified using RNeasy MinElute spin columns (see flowchart " miRNeasy FFPE procedure"). Since RNA is eluted in a volume of just 14–30 µl, smaller reaction volumes are possible in downstream applications.
See figures
Applications
The miRNeasy FFPE Kit allows purification of miRNA with total RNA for use in a variety of applications, such as quantitative, real-time RT-PCR.
Supporting data and figures
Efficient RNA purification from FFPE tissues.
Total RNA including miRNA was purified from the indicated rat tissues using either the miRNeasy FFPE Kit or phenol-chloroform extraction. Purified RNA was used as a template in quantitative, real-time RT-PCR assays for the miRNA miR-16. Results showed that for lung and liver tissues, CT values were lower after purification using the miRNeasy FFPE Kit, indicating that higher amounts of miRNA were purified than when using phenol-chloroform extraction. For kidney tissue, CT values were similar for both methods. CT values were similar or lower after purification using the miRNeasy FFPE Kit for all tissue types tested.
Specifications
| Features | Specifications |
|---|---|
| applications | PCR, qPRC, real-time RT-PCR, microArray |
| elutionvolume | 14-30 µl |
| purificationoftotalrnamirnapolyamrnadnaorprotein | miRNA, total RNA |
| sampleamount | up to 4 sections, each with a thickness up to 10 µm and a surface area up to 250mm^2 (automated protocol on QIAcube: up to 2 sections) |
| processing | Manual (protocol for automated processing on QIAcube available) |
| mainsampletype | FFPE tissue samples |
| format | Spin column |
| technology | Silica technology |
| yield | Varies |
Resources
Brochures & Guides en (5)
Safety Data Sheets (1)
Scientific Posters (1)
Quick-Start Protocols (2)
Application Notes (1)
Brochures & Guides (1)
Kit Handbooks (1)
Additional Resources (1)
miRNA Research (1)
Certificates of Analysis (1)