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AllPrep DNA/RNA FFPE Kit (50)
Cat. No. / ID: 80234
50 RNeasy MinElute Spin Columns, 50 QIAamp MinElute Spin Columns, Collection Tubes, RNase-Free Reagents, and Buffers
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The AllPrep DNA/RNA 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 for your AllPrep DNA/RNA FFPE Kit (50) trial kit.
Features
- Maximum output with minimal sample consumption
- Releases DNA/RNA without compromising integrity
- Effectively separates RNA and DNA
- Comprehensive DNA and RNA analysis of the same FFPE sample
Product Details
For reliable comparison of genomic and transcriptomic data, purification of DNA and RNA from the same sample is essential. The AllPrep DNA/RNA FFPE Kit uses a patent-pending solubilization method to purify DNA and RNA from formalin-fixed, paraffin-embedded (FFPE) tissue sections. Purified analytes are suitable for use in applications such as real-time PCR and Pyrosequencing. The kit can be automated on the QIAcube Connect.
Performance
DNA and RNA purified using the AllPrep DNA/RNA FFPE Kit are of comparable quality to DNA and RNA purified using the QIAamp FFPE Tissue Kit and RNeasy FFPE Kit/miRNeasy FFPE, respectively (see figure " Purification of DNA and RNA from FFPE samples"). The purified nucleic acids are therefore suitable for downstream applications such as Pyrosequencing or real-time PCR and RT-PCR (see figure " Reliable amplification of DNA and RNA from FFPE samples" and " Array analysis following preamplification of cDNA targets").
See figures
Principle
The AllPrep DNA/RNA FFPE Kit is specially designed for simultaneous purification of genomic DNA and total RNA from FFPE tissue sections. Pure DNA and RNA are obtained from the entire sample, in contrast to other procedures where the biological sample is divided into two before being processed separately. Simply dividing a sample in half for separate DNA and RNA purification procedures results in the purification of DNA and RNA from different populations of cells, which may differ in their properties. Purification of DNA and RNA from the same sample also helps to prevent waste, since FFPE samples are precious, often difficult to retrieve, and limited in amount.
Due to fixation and embedding conditions, nucleic acids in FFPE samples are usually heavily fragmented and are often of a lower molecular weight than those obtained from fresh or frozen samples. A major obstacle to isolating DNA and RNA from the same FFPE sample is that fragmented DNA is short and can be partly single-stranded and therefore more closely resembles RNA than intact DNA. This property of fragmented DNA makes physical separation of DNA and RNA difficult. The AllPrep DNA/RNA FFPE Kit uses a patent-pending solubilization method to differentially release DNA and RNA from a single FFPE sample.
Nucleic acids in FFPE samples are also chemically modified by formaldehyde. Although formaldehyde modification cannot be detected in standard quality control assays, such as gel electrophoresis or lab-on-a-chip analysis, it does strongly interfere with enzymatic analyses. The AllPrep DNA/RNA FFPE Kit is optimized to reverse as much formaldehyde modification as possible without further DNA and RNA degradation.
Procedure
A simple workflow allows the purification of high-quality DNA and RNA from the same FFPE tissue section sample (see flowchart " AllPrep DNA/RNA FFPE procedure"). The AllPrep DNA/RNA FFPE Kit uses a patent-pending solubilization method to differentially release DNA and RNA from a single FFPE sample. With this method, FFPE samples are incubated in an optimized lysis buffer, which results in the release of RNA and precipitation of DNA. After centrifugation, the RNA-containing supernatant and DNA-containing pellet are then processed separately to purify RNA and DNA. Further incubations partially reverse crosslinking, and RNA or DNA is then purified using an RNeasy MinElute spin column or QIAamp MinElute spin column. For purified RNA, an on-column DNase treatment efficiently removes any contaminating DNA. Depending on the RNA binding conditions, small RNAs such as miRNA are either absent or present in the purified RNA. For purified DNA, an on-column RNase treatment is optional, as RNA contamination is minimal due to the separation of DNA and RNA prior to spin column processing.
See figures
Applications
The AllPrep DNA/RNA FFPE Kit is optimized to reverse as much formaldehyde modification as possible without further DNA and RNA degradation; however nucleic acids purified from FFPE samples should not be used in downstream applications that require high-molecular-weight DNA or full-length RNA. Some applications may require modifications to allow the use of fragmented nucleic acids (e.g., designing small amplicons for PCR and RT-PCR). For cDNA synthesis, gene-specific primers should be used instead of oligo-dT primers. If it is not possible to use gene-specific primers, random primers should be used.
Supporting data and figures
Purification of DNA and RNA from FFPE samples.
[A] Genomic DNA was purified from various FFPE rat tissues that were stored at room temperature for the times indicated. Purification was performed using either the AllPrep DNA/RNA FFPE Kit or, as a control, the QIAamp DNA FFPE Tissue Kit (a dedicated kit for DNA purification from FFPE samples) both including RNase digestion. DNA yields from 20 μm sections of each sample were determined by absorbance measurement. [B] RNA was purified from various FFPE rat tissues that were stored at room temperature for the times indicated. Purification was performed using either the AllPrep DNA/RNA FFPE Kit or, as a control, the RNeasy FFPE Kit (a dedicated kit for RNA purification from FFPE samples). RNA yields from 10 μm sections of each sample were determined by absorbance measurement. The AllPrep Kit performed just as well as the dedicated DNA/RNA purification kits in recovering DNA/RNA from FFPE samples.
Specifications
| Features | Specifications |
|---|---|
| Applications | PCR, qPCR, real-time RT-PCR, microarray |
| Elution volume | RNA: 14-30µl ; DNA: 30-100µl |
| Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein | RNA (miRNA) and DNA |
| Processing | Manual (centrifugation) |
| Format | Spin column |
| Number of preps per run | 50 |
| Sample amount | max. 4*10 µm sections or 2*20 µm sections |
| Technology | Silica technology |
| Time per run or per prep | 6h for 10 samples, including sectioning |
| Yield | Varies |
| Main sample type | FFPE tissue samples |
Resources
Brochures & Guides (3)
Quick-Start Protocols (2)
Scientific Posters (1)
Kit Handbooks (1)
Safety Data Sheets (1)
Supplementary Protocols (2)
Additional Resources (1)
Certificates of Analysis (1)
FAQ
What is the composition of Buffer FRN?
What is the composition of Buffer PKD?
How much DNA/RNA can be isolated with the AllPrep DNA/RNA FFPE Kit?
How long can I store my purified DNA/RNA isolated with the AllPrep DNA/RNA FFPE kit?
Is the yield and quality of RNA and DNA purified with the AllPrep DNA/RNA FFPE Kit comparable to yield and quality of nucleic acids purified separately with other methods.
Which deparaffinization method can I use with the AllPrep DNA/RNA FFPE Kit?
Is it also possible to isolate miRNA with the AllPrep DNA/RNA FFPE Kit?
How much sample material can I use for DNA/RNA purification with the AllPrep DNA/RNA FFPE Kit?
What is Tissue-Tek O.C.T., and what is it used for?